anti rbd monoclonal antibody  (Sino Biological)


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    Name:
    SARS CoV SARS CoV 2 Spike antibody Chimeric MAb
    Description:
    It is a chimeric monoclonal antibody combining the constant domains of the human IgG1 molecule with mouse variable regions The variable region was obtained from a mouse immunized with purified recombinant SARS CoV Spike RBD Protein The antibody was produced using recombinant antibody technology
    Catalog Number:
    40150-d001
    Product Aliases:
    Anti-coronavirus s1 Antibody, Anti-coronavirus s2 Antibody, Anti-coronavirus spike Antibody, Anti-cov spike Antibody, Anti-ncov RBD Antibody, Anti-ncov s1 Antibody, Anti-ncov s2 Antibody, Anti-ncov spike Antibody, Anti-novel coronavirus RBD Antibody, Anti-novel coronavirus s1 Antibody, Anti-novel coronavirus s2 Antibody, Anti-novel coronavirus spike Antibody, Anti-RBD Antibody, Anti-S1 Antibody, Anti-s2 Antibody, Anti-Spike RBD Antibody
    Price:
    None
    Applications:
    ELISA
    Host:
    Chimera
    Immunogen:
    Recombinant SARS-CoV Spike RBD Protein (Catalog#40150-V08B2)
    Category:
    Primary Antibody
    Antibody Type:
    MAb
    Isotype:
    mouse (variable region) / human (kappa / IgG1 constant) chimeric antibody
    Reactivity:
    SARS
    Buy from Supplier


    Structured Review

    Sino Biological anti rbd monoclonal antibody
    Detection of <t>SARS-CoV-2</t> spike (S) protein expression and localization. (A) Schematic illustration of the SARS-CoV-2 full-length spike (S-FL) and mutant S variants. The <t>RBD</t> (receptor binding domain) is in subunit S1; the FP (fusion peptide), HR1 (heptad repeat 1), HR2 (heptad repeat 2), TM (transmembrane domain), and CT (cytoplasmic tail) are in subunit S2. The endoplasmic reticulum retrieval signals (“KxHxx” motif) in the CT domain of S-FL were destroyed in S-Mut protein. The C-terminal 19 amino acids were lacking in S-C19del. (B) Detection of SARS-CoV-2 S expression in HKE293T cells by Western blot using the anti-RBD monoclonal antibody. Cells were transfected with pS-FL, pS-Mut, and pS-C19del plasmids or with an empty vector. (C) Detection of SARS-CoV-2 S subcellular localization in HKE293T cells by confocal microscopy. Cells were grown on glass coverslips for 24 h preceding transfection of plasmids encoding S protein variants. The cells were harvested and labeled with the corresponding antibodies. Calreticulin, ER marker. Nuclei were counterstained with DAPI. Bar = 20 μm.
    It is a chimeric monoclonal antibody combining the constant domains of the human IgG1 molecule with mouse variable regions The variable region was obtained from a mouse immunized with purified recombinant SARS CoV Spike RBD Protein The antibody was produced using recombinant antibody technology
    https://www.bioz.com/result/anti rbd monoclonal antibody/product/Sino Biological
    Average 95 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    anti rbd monoclonal antibody - by Bioz Stars, 2021-02
    95/100 stars

    Images

    1) Product Images from "Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2"

    Article Title: Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2

    Journal: Genes & Diseases

    doi: 10.1016/j.gendis.2020.07.006

    Detection of SARS-CoV-2 spike (S) protein expression and localization. (A) Schematic illustration of the SARS-CoV-2 full-length spike (S-FL) and mutant S variants. The RBD (receptor binding domain) is in subunit S1; the FP (fusion peptide), HR1 (heptad repeat 1), HR2 (heptad repeat 2), TM (transmembrane domain), and CT (cytoplasmic tail) are in subunit S2. The endoplasmic reticulum retrieval signals (“KxHxx” motif) in the CT domain of S-FL were destroyed in S-Mut protein. The C-terminal 19 amino acids were lacking in S-C19del. (B) Detection of SARS-CoV-2 S expression in HKE293T cells by Western blot using the anti-RBD monoclonal antibody. Cells were transfected with pS-FL, pS-Mut, and pS-C19del plasmids or with an empty vector. (C) Detection of SARS-CoV-2 S subcellular localization in HKE293T cells by confocal microscopy. Cells were grown on glass coverslips for 24 h preceding transfection of plasmids encoding S protein variants. The cells were harvested and labeled with the corresponding antibodies. Calreticulin, ER marker. Nuclei were counterstained with DAPI. Bar = 20 μm.
    Figure Legend Snippet: Detection of SARS-CoV-2 spike (S) protein expression and localization. (A) Schematic illustration of the SARS-CoV-2 full-length spike (S-FL) and mutant S variants. The RBD (receptor binding domain) is in subunit S1; the FP (fusion peptide), HR1 (heptad repeat 1), HR2 (heptad repeat 2), TM (transmembrane domain), and CT (cytoplasmic tail) are in subunit S2. The endoplasmic reticulum retrieval signals (“KxHxx” motif) in the CT domain of S-FL were destroyed in S-Mut protein. The C-terminal 19 amino acids were lacking in S-C19del. (B) Detection of SARS-CoV-2 S expression in HKE293T cells by Western blot using the anti-RBD monoclonal antibody. Cells were transfected with pS-FL, pS-Mut, and pS-C19del plasmids or with an empty vector. (C) Detection of SARS-CoV-2 S subcellular localization in HKE293T cells by confocal microscopy. Cells were grown on glass coverslips for 24 h preceding transfection of plasmids encoding S protein variants. The cells were harvested and labeled with the corresponding antibodies. Calreticulin, ER marker. Nuclei were counterstained with DAPI. Bar = 20 μm.

    Techniques Used: Expressing, Mutagenesis, Binding Assay, Western Blot, Transfection, Plasmid Preparation, Confocal Microscopy, Labeling, Marker

    2) Product Images from "A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)"

    Article Title: A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)

    Journal: Biosensors & Bioelectronics

    doi: 10.1016/j.bios.2020.112715

    Sensitivity and specificity of the ACE2-based LFA a) Results of ACE2-based LFA for the detection sensitivity of SARS-CoV-2 S1 antigen. Serially diluted antigen concentrates (concentration range: 500 ng/mL to 5 ng/mL) were tested by ACE2-based LFA. After 20 min, the LFA strips were photographed with a smartphone. Moreover, the intensity of the test and control lines was converted to a peak histogram by an image analyzer. b) Results of the comparative analysis for the detection selectivity: positive control – SARS-CoV-2 S1, negative control – SARS-CoV S1, MERS S1, and buffer solution. Using three different concentrates (1 μg/mL, 200 ng/mL, and 50 ng/mL) of the antigen sample, the detection performance of ACE2-based LFA was demonstrated. c) Bar graph of peak intensities for test lines. After 20 min for the sample flow, the intensity of the test lines was measured by a portable line analyzer. Inset) the detection intensity for the 5 ng antigen per reaction of each control. Limit of detection (LOD) was determined by the mean value of negative controls plus three times the standard deviation. P-values: ns > 0.05, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.
    Figure Legend Snippet: Sensitivity and specificity of the ACE2-based LFA a) Results of ACE2-based LFA for the detection sensitivity of SARS-CoV-2 S1 antigen. Serially diluted antigen concentrates (concentration range: 500 ng/mL to 5 ng/mL) were tested by ACE2-based LFA. After 20 min, the LFA strips were photographed with a smartphone. Moreover, the intensity of the test and control lines was converted to a peak histogram by an image analyzer. b) Results of the comparative analysis for the detection selectivity: positive control – SARS-CoV-2 S1, negative control – SARS-CoV S1, MERS S1, and buffer solution. Using three different concentrates (1 μg/mL, 200 ng/mL, and 50 ng/mL) of the antigen sample, the detection performance of ACE2-based LFA was demonstrated. c) Bar graph of peak intensities for test lines. After 20 min for the sample flow, the intensity of the test lines was measured by a portable line analyzer. Inset) the detection intensity for the 5 ng antigen per reaction of each control. Limit of detection (LOD) was determined by the mean value of negative controls plus three times the standard deviation. P-values: ns > 0.05, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.

    Techniques Used: Concentration Assay, Positive Control, Negative Control, Standard Deviation

    Laboratory confirmation of ACE2-based LFIA using clinical samples a) Schematic diagram of COVID-19 test using ACE2-based LFIA. A nasopharyngeal swab from the COVID-19 patient is placed into the UTM. 50 μL of UTM containing the SARS-CoV-2 is mixed with running buffer in a 1:1 (v/v) ratio, and 100 μL of mixed solution is loaded into the LFIA device. After 20 min, the line intensity of the LFIA strip is semi-quantified by the portable analyzer. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 10 8 copies/mL to 5.35 × 10 6 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (I L : line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control. c) Bar graph of intensities for test lines measured by the portable analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (0 copies/mL of SARS-CoV-2) plus three times the standard deviation. d) Laboratory confirmation of ACE2-based LFIA compared to the RT-qPCR using clinical samples. i) Nasopharyngeal swab samples of COVID-19 patients (n = 4) and healthy subjects (n = 4) were tested both ACE2-based LFIA and RT-qPCR. Sensitivity was determined by the number of true positive samples divided by the number of positive samples tested. Moreover, specificity was determined by the number of true negative samples divide by the number of negative samples tested. ii) RT-qPCR results for the detection of the SARS-CoV-2 specific gene (Env gene). C t value and their correspondent viral load in the clinical samples were evaluated. e) Results of ACE2-based LFIA on laboratory confirmation using clinical COVID-19 patient samples. Twenty minutes after sample loading, the test line intensities of the LFIA strips were measured with a portable line analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (healthy control) plus three times the standard deviation.
    Figure Legend Snippet: Laboratory confirmation of ACE2-based LFIA using clinical samples a) Schematic diagram of COVID-19 test using ACE2-based LFIA. A nasopharyngeal swab from the COVID-19 patient is placed into the UTM. 50 μL of UTM containing the SARS-CoV-2 is mixed with running buffer in a 1:1 (v/v) ratio, and 100 μL of mixed solution is loaded into the LFIA device. After 20 min, the line intensity of the LFIA strip is semi-quantified by the portable analyzer. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 10 8 copies/mL to 5.35 × 10 6 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (I L : line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control. c) Bar graph of intensities for test lines measured by the portable analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (0 copies/mL of SARS-CoV-2) plus three times the standard deviation. d) Laboratory confirmation of ACE2-based LFIA compared to the RT-qPCR using clinical samples. i) Nasopharyngeal swab samples of COVID-19 patients (n = 4) and healthy subjects (n = 4) were tested both ACE2-based LFIA and RT-qPCR. Sensitivity was determined by the number of true positive samples divided by the number of positive samples tested. Moreover, specificity was determined by the number of true negative samples divide by the number of negative samples tested. ii) RT-qPCR results for the detection of the SARS-CoV-2 specific gene (Env gene). C t value and their correspondent viral load in the clinical samples were evaluated. e) Results of ACE2-based LFIA on laboratory confirmation using clinical COVID-19 patient samples. Twenty minutes after sample loading, the test line intensities of the LFIA strips were measured with a portable line analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (healthy control) plus three times the standard deviation.

    Techniques Used: Stripping Membranes, Cell Culture, Concentration Assay, Negative Control, Standard Deviation, Quantitative RT-PCR

    Cellular receptor (ACE2)-based LFIA. a) Schematic of ACE2 receptor recognition by SARS-CoV-2. ACE2, a type 1 membrane protein expressed in the lung, heart, kidney, and intestine, is the cellular receptor for SARS-CoV-2. b) Schematic of an ACE2-based LFIA consisting of a sample pad, conjugate pad, nitrocellulose membrane, and absorbent pad. The test line placed on the nitrocellulose membrane contains ACE2 for detection of the SARS-CoV-2 spike antigen. Anti-IgG antibody is used in the control line. The proposed LFIA can achieve sensitive and selective detection of SARS-CoV-2 spike antigen within 20 min.
    Figure Legend Snippet: Cellular receptor (ACE2)-based LFIA. a) Schematic of ACE2 receptor recognition by SARS-CoV-2. ACE2, a type 1 membrane protein expressed in the lung, heart, kidney, and intestine, is the cellular receptor for SARS-CoV-2. b) Schematic of an ACE2-based LFIA consisting of a sample pad, conjugate pad, nitrocellulose membrane, and absorbent pad. The test line placed on the nitrocellulose membrane contains ACE2 for detection of the SARS-CoV-2 spike antigen. Anti-IgG antibody is used in the control line. The proposed LFIA can achieve sensitive and selective detection of SARS-CoV-2 spike antigen within 20 min.

    Techniques Used:

    Indirect ELISA results from spike antigens of three different coronaviruses (SARS-CoV S1, SARS-CoV-2 S1, and MERS S1). a) The interactions between these S1 antigens and ACE2 were examined with serially diluted samples (concentration range: 200 to 0.05 ng/mL). In addition, three different antibodies, CR3022 (black) (b); F26G19 (red) (c); and S1-mAb (orange) (d), were tested for their interaction with spike antigens, using the same concentration range. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
    Figure Legend Snippet: Indirect ELISA results from spike antigens of three different coronaviruses (SARS-CoV S1, SARS-CoV-2 S1, and MERS S1). a) The interactions between these S1 antigens and ACE2 were examined with serially diluted samples (concentration range: 200 to 0.05 ng/mL). In addition, three different antibodies, CR3022 (black) (b); F26G19 (red) (c); and S1-mAb (orange) (d), were tested for their interaction with spike antigens, using the same concentration range. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

    Techniques Used: Indirect ELISA, Concentration Assay

    Identification of the sandwich pair for detection of SARS-CoV-2 spike antigen. a) Schematic diagram of LFIA using ACE2 as the capture probe and sandwich analysis results obtained from paired antibodies (CR3022, F26G19, and S1mAb). SARS-CoV-2 S1 antigen (50 ng) was used as a positive control, and buffer containing no S1 antigen was used as a negative control. After 20 min, the strips were captured by a smartphone, and their peak intensities were analyzed. b) Schematic diagram of LFIA, using antibodies as the capture probe, and their sandwich analysis results. c) Peak intensities of capture probe (P C )–detection probe (P D ) pairs. A total of 12 pairs of positive controls (50 ng S1 antigen) were tested, and their intensities were analyzed. Peak intensity was calculated by subtracting the background intensity of the strip from the average intensities of the dots.
    Figure Legend Snippet: Identification of the sandwich pair for detection of SARS-CoV-2 spike antigen. a) Schematic diagram of LFIA using ACE2 as the capture probe and sandwich analysis results obtained from paired antibodies (CR3022, F26G19, and S1mAb). SARS-CoV-2 S1 antigen (50 ng) was used as a positive control, and buffer containing no S1 antigen was used as a negative control. After 20 min, the strips were captured by a smartphone, and their peak intensities were analyzed. b) Schematic diagram of LFIA, using antibodies as the capture probe, and their sandwich analysis results. c) Peak intensities of capture probe (P C )–detection probe (P D ) pairs. A total of 12 pairs of positive controls (50 ng S1 antigen) were tested, and their intensities were analyzed. Peak intensity was calculated by subtracting the background intensity of the strip from the average intensities of the dots.

    Techniques Used: Positive Control, Negative Control, Stripping Membranes

    Biolayer interferometry (BLI) results of ACE2, CR3022, F26G19, and S1-mAb against the SARS-CoV-2 S1 antigen. Dotted lines represent the response curves of BLI measurement, and solid lines represent the fitting curves based on a 1:1 binding model. Binding kinetics were measured for four different concentrations of the S1 antigen.
    Figure Legend Snippet: Biolayer interferometry (BLI) results of ACE2, CR3022, F26G19, and S1-mAb against the SARS-CoV-2 S1 antigen. Dotted lines represent the response curves of BLI measurement, and solid lines represent the fitting curves based on a 1:1 binding model. Binding kinetics were measured for four different concentrations of the S1 antigen.

    Techniques Used: Binding Assay

    3) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    4) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    5) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    6) Product Images from "CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells"

    Article Title: CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells

    Journal: Signal Transduction and Targeted Therapy

    doi: 10.1038/s41392-020-00426-x

    CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p
    Figure Legend Snippet: CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p

    Techniques Used: Infection, Co-Immunoprecipitation Assay, Microscopy, Immunofluorescence, Multicolor Immunofluorescence Staining, Real-time Polymerase Chain Reaction

    7) Product Images from "SARS-CoV-2 invades host cells via a novel route: CD147-spike protein"

    Article Title: SARS-CoV-2 invades host cells via a novel route: CD147-spike protein

    Journal: bioRxiv

    doi: 10.1101/2020.03.14.988345

    The localization of CD147 and SP observed by immune-electron microscope. (A) The SARS-CoV-2 infected Vero E6 cell with visible SARS-CoV-2 virion (black arrow). ( B and C ) The localization of CD147 (10nm-gold colloid, yellow arrows) and SP (20nm-gold colloid, red arrows) in viral inclusion bodies of SARS-CoV-2 infected Vero E6 cell. Scale bar = 200nm.
    Figure Legend Snippet: The localization of CD147 and SP observed by immune-electron microscope. (A) The SARS-CoV-2 infected Vero E6 cell with visible SARS-CoV-2 virion (black arrow). ( B and C ) The localization of CD147 (10nm-gold colloid, yellow arrows) and SP (20nm-gold colloid, red arrows) in viral inclusion bodies of SARS-CoV-2 infected Vero E6 cell. Scale bar = 200nm.

    Techniques Used: Microscopy, Infection

    Identification of interaction between CD147 and SP. (A) The interaction of CD147 and SP detected by SPR assay, KD = 1.85×10 −7 M. (B) The interaction of CD147 and SP detected by Co-IP assay. Anti-CD147 antibody and anti-SARS-CoV-2 Spike antibody were used for antibody immobilization for Co-IP. The mIgG and rIgG were selected as negative control. (C) The interaction of CD147 and SP detected by ELISA. (D) The ability of Meplazumab to compete with SP for CD147 binding performed by competitive inhibition ELISA.
    Figure Legend Snippet: Identification of interaction between CD147 and SP. (A) The interaction of CD147 and SP detected by SPR assay, KD = 1.85×10 −7 M. (B) The interaction of CD147 and SP detected by Co-IP assay. Anti-CD147 antibody and anti-SARS-CoV-2 Spike antibody were used for antibody immobilization for Co-IP. The mIgG and rIgG were selected as negative control. (C) The interaction of CD147 and SP detected by ELISA. (D) The ability of Meplazumab to compete with SP for CD147 binding performed by competitive inhibition ELISA.

    Techniques Used: SPR Assay, Co-Immunoprecipitation Assay, Negative Control, Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition

    8) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    9) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    10) Product Images from "Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens"

    Article Title: Development of immunohistochemistry and in situ hybridisation for the detection of SARS-CoV and SARS-CoV-2 in formalin-fixed paraffin-embedded specimens

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-78949-0

    Immunohistochemistry labelling of FFPE cells expressing SARS-CoV, SARS-CoV-2 and MERS spike proteins. Immunodetection performed using SARS-CoV spike rabbit monoclonal antibody on producer cells for SARS-CoV (a) , SARS-CoV-2 (b) and MERS-CoV pseudotype virus (c) and non-transfected cells (d) . Scale bars, 20 µm.
    Figure Legend Snippet: Immunohistochemistry labelling of FFPE cells expressing SARS-CoV, SARS-CoV-2 and MERS spike proteins. Immunodetection performed using SARS-CoV spike rabbit monoclonal antibody on producer cells for SARS-CoV (a) , SARS-CoV-2 (b) and MERS-CoV pseudotype virus (c) and non-transfected cells (d) . Scale bars, 20 µm.

    Techniques Used: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Expressing, Immunodetection, Transfection

    Immunohistochemistry and in situ hybridisation detection of SARS-CoV-2 and RNA on infected ferret tissues. Detection of spike protein (a) , nucleoprotein (b) and dsRNA antigens (c) and spike RNA (d) labelling. Tissue shrinkage artefact with ISH pre-treatment (d) . Scale bars, 20 µm.
    Figure Legend Snippet: Immunohistochemistry and in situ hybridisation detection of SARS-CoV-2 and RNA on infected ferret tissues. Detection of spike protein (a) , nucleoprotein (b) and dsRNA antigens (c) and spike RNA (d) labelling. Tissue shrinkage artefact with ISH pre-treatment (d) . Scale bars, 20 µm.

    Techniques Used: Immunohistochemistry, In Situ, Hybridization, Infection, In Situ Hybridization

    Immunohistochemical labelling of FFPE SARS-CoV and SARS-CoV-2 infected cells and uninfected cells. Immunodetection performed using SARS-CoV spike rabbit monoclonal antibody (a–c) , SARS-CoV nucleoprotein rabbit polyclonal antibody (d–f) and double-stranded RNA (dsRNA) rabbit monoclonal antibody (g–i) . Scale bars, 20 µm.
    Figure Legend Snippet: Immunohistochemical labelling of FFPE SARS-CoV and SARS-CoV-2 infected cells and uninfected cells. Immunodetection performed using SARS-CoV spike rabbit monoclonal antibody (a–c) , SARS-CoV nucleoprotein rabbit polyclonal antibody (d–f) and double-stranded RNA (dsRNA) rabbit monoclonal antibody (g–i) . Scale bars, 20 µm.

    Techniques Used: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Infection, Immunodetection

    In situ hybridisation (ISH) of FFPE cells infected with SARS-CoV and SARS-CoV-2 using RNAScope ® . ISH performed using RNA probes designed specific to SARS-CoV-2 spike RNA. SARS-CoV (a) and SARS-CoV-2 infected cells (b) , uninfected cells (c) . Scale bars, 20 µm.
    Figure Legend Snippet: In situ hybridisation (ISH) of FFPE cells infected with SARS-CoV and SARS-CoV-2 using RNAScope ® . ISH performed using RNA probes designed specific to SARS-CoV-2 spike RNA. SARS-CoV (a) and SARS-CoV-2 infected cells (b) , uninfected cells (c) . Scale bars, 20 µm.

    Techniques Used: In Situ, Hybridization, In Situ Hybridization, Formalin-fixed Paraffin-Embedded, Infection

    11) Product Images from "A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)"

    Article Title: A novel rapid detection for SARS-CoV-2 spike 1 antigens using human angiotensin converting enzyme 2 (ACE2)

    Journal: Biosensors & Bioelectronics

    doi: 10.1016/j.bios.2020.112715

    Laboratory confirmation of ACE2-based LFIA using clinical samples a) Schematic diagram of COVID-19 test using ACE2-based LFIA. A nasopharyngeal swab from the COVID-19 patient is placed into the UTM. 50 μL of UTM containing the SARS-CoV-2 is mixed with running buffer in a 1:1 (v/v) ratio, and 100 μL of mixed solution is loaded into the LFIA device. After 20 min, the line intensity of the LFIA strip is semi-quantified by the portable analyzer. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 10 8 copies/mL to 5.35 × 10 6 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (I L : line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control. c) Bar graph of intensities for test lines measured by the portable analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (0 copies/mL of SARS-CoV-2) plus three times the standard deviation. d) Laboratory confirmation of ACE2-based LFIA compared to the RT-qPCR using clinical samples. i) Nasopharyngeal swab samples of COVID-19 patients (n = 4) and healthy subjects (n = 4) were tested both ACE2-based LFIA and RT-qPCR. Sensitivity was determined by the number of true positive samples divided by the number of positive samples tested. Moreover, specificity was determined by the number of true negative samples divide by the number of negative samples tested. ii) RT-qPCR results for the detection of the SARS-CoV-2 specific gene (Env gene). C t value and their correspondent viral load in the clinical samples were evaluated. e) Results of ACE2-based LFIA on laboratory confirmation using clinical COVID-19 patient samples. Twenty minutes after sample loading, the test line intensities of the LFIA strips were measured with a portable line analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (healthy control) plus three times the standard deviation.
    Figure Legend Snippet: Laboratory confirmation of ACE2-based LFIA using clinical samples a) Schematic diagram of COVID-19 test using ACE2-based LFIA. A nasopharyngeal swab from the COVID-19 patient is placed into the UTM. 50 μL of UTM containing the SARS-CoV-2 is mixed with running buffer in a 1:1 (v/v) ratio, and 100 μL of mixed solution is loaded into the LFIA device. After 20 min, the line intensity of the LFIA strip is semi-quantified by the portable analyzer. b) Results of ACE2-based LFA for the detection sensitivity of cultured SARS-CoV-2. Serially diluted virus concentrates (concentration range: 1.07 × 10 8 copies/mL to 5.35 × 10 6 copies/mL) were tested. After 20 min, the LFIA strip was taken with a smartphone and scanned with an image analyzer. The line intensities of the test and control lines were converted to peak histograms. Also, the intensity of the test lines was measured by a portable line analyzer (I L : line intensity of test line). Furthermore, human coronavirus (OC43) was tested as a negative control. c) Bar graph of intensities for test lines measured by the portable analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (0 copies/mL of SARS-CoV-2) plus three times the standard deviation. d) Laboratory confirmation of ACE2-based LFIA compared to the RT-qPCR using clinical samples. i) Nasopharyngeal swab samples of COVID-19 patients (n = 4) and healthy subjects (n = 4) were tested both ACE2-based LFIA and RT-qPCR. Sensitivity was determined by the number of true positive samples divided by the number of positive samples tested. Moreover, specificity was determined by the number of true negative samples divide by the number of negative samples tested. ii) RT-qPCR results for the detection of the SARS-CoV-2 specific gene (Env gene). C t value and their correspondent viral load in the clinical samples were evaluated. e) Results of ACE2-based LFIA on laboratory confirmation using clinical COVID-19 patient samples. Twenty minutes after sample loading, the test line intensities of the LFIA strips were measured with a portable line analyzer. The limit of detection (LOD) was determined by the mean value of negative controls (healthy control) plus three times the standard deviation.

    Techniques Used: Stripping Membranes, Cell Culture, Concentration Assay, Negative Control, Standard Deviation, Quantitative RT-PCR

    12) Product Images from "Clinical and pathological findings of SARS-CoV-2 infection and concurrent IgA nephropathy: a case report"

    Article Title: Clinical and pathological findings of SARS-CoV-2 infection and concurrent IgA nephropathy: a case report

    Journal: BMC Nephrology

    doi: 10.1186/s12882-020-02163-3

    Renal biopsy findings. a Glomerulus with a fibrocelluar crescent, adjacent acute tubular injury, as well as associated tubular atrophy/interstitial inflammation (PAS stain; original magnification × 200); b Cells debris within the proximal tubular lumen (PAS stain; original magnification × 400); c direct immunofluorescence staining with IgA (original magnification × 400); d Ultrastructure examination reveals mesangial electron-dense deposits (transmission electron microscopy; original magnification × 2500); e Negative immunohistochemistry staining for the S1 spike protein of SARS-CoV-2 (original magnification × 200). PAS: Periodic Acid-Schiff; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2
    Figure Legend Snippet: Renal biopsy findings. a Glomerulus with a fibrocelluar crescent, adjacent acute tubular injury, as well as associated tubular atrophy/interstitial inflammation (PAS stain; original magnification × 200); b Cells debris within the proximal tubular lumen (PAS stain; original magnification × 400); c direct immunofluorescence staining with IgA (original magnification × 400); d Ultrastructure examination reveals mesangial electron-dense deposits (transmission electron microscopy; original magnification × 2500); e Negative immunohistochemistry staining for the S1 spike protein of SARS-CoV-2 (original magnification × 200). PAS: Periodic Acid-Schiff; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2

    Techniques Used: Staining, Immunofluorescence, Transmission Assay, Electron Microscopy, Immunohistochemistry

    13) Product Images from "Process Development and Scale-up Optimization of the SARS-CoV-2 Receptor Binding Domain-Based Vaccine Candidate, RBD219-N1C1"

    Article Title: Process Development and Scale-up Optimization of the SARS-CoV-2 Receptor Binding Domain-Based Vaccine Candidate, RBD219-N1C1

    Journal: bioRxiv

    doi: 10.1101/2020.12.30.424829

    Time point SDS-PAGE analysis of pre- and post-induction fermentation samples of the lockdown process (Run 5). PI: pre-induction; D1, D2, D3: Day 1-3 after induction. The arrow shows RBD219-N1C1 in fermentation supernatant after induction.
    Figure Legend Snippet: Time point SDS-PAGE analysis of pre- and post-induction fermentation samples of the lockdown process (Run 5). PI: pre-induction; D1, D2, D3: Day 1-3 after induction. The arrow shows RBD219-N1C1 in fermentation supernatant after induction.

    Techniques Used: SDS Page

    Impurity evaluation of the purified RBD219-N1C1 proteins from three processes. (A-B) Unpurified (FS) and purified RBD219-N1C1 in reduced SDS-PAGE with Coomassie Blue stain (A) and with Western blot using anti- P. pastoris HCP antibody (B). (C) Measured P. pastoris HCP content by quantitative ELISA and (D) endotoxin levels are shown.
    Figure Legend Snippet: Impurity evaluation of the purified RBD219-N1C1 proteins from three processes. (A-B) Unpurified (FS) and purified RBD219-N1C1 in reduced SDS-PAGE with Coomassie Blue stain (A) and with Western blot using anti- P. pastoris HCP antibody (B). (C) Measured P. pastoris HCP content by quantitative ELISA and (D) endotoxin levels are shown.

    Techniques Used: Purification, SDS Page, Staining, Western Blot, Enzyme-linked Immunosorbent Assay

    Characterization of purified RBD219-N1C1 proteins from three processes. (A-B) Purified proteins are analyzed by SDS-PAGE gel with Coomassie Blue stain (A) and Western blot with a monoclonal SARS-CoV-2 Spike antibody (B). (C-D) Size and aggregates evaluation by SEC-HPLC (C) and the radius and size in solution measured by Dynamic Light Scattering (D). (D) Averages ± SD are shown from four independent measurements.
    Figure Legend Snippet: Characterization of purified RBD219-N1C1 proteins from three processes. (A-B) Purified proteins are analyzed by SDS-PAGE gel with Coomassie Blue stain (A) and Western blot with a monoclonal SARS-CoV-2 Spike antibody (B). (C-D) Size and aggregates evaluation by SEC-HPLC (C) and the radius and size in solution measured by Dynamic Light Scattering (D). (D) Averages ± SD are shown from four independent measurements.

    Techniques Used: Purification, SDS Page, Staining, Western Blot, High Performance Liquid Chromatography

    Binding ability of the purified RBD219-N1C1 from three processes to a recombinant human ACE2 receptor
    Figure Legend Snippet: Binding ability of the purified RBD219-N1C1 from three processes to a recombinant human ACE2 receptor

    Techniques Used: Binding Assay, Purification, Recombinant

    14) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    15) Product Images from "Identification of Immunohistochemical Reagents for In Situ Protein Expression Analysis of Coronavirus-associated Changes in Human Tissues"

    Article Title: Identification of Immunohistochemical Reagents for In Situ Protein Expression Analysis of Coronavirus-associated Changes in Human Tissues

    Journal: Applied Immunohistochemistry & Molecular Morphology

    doi: 10.1097/PAI.0000000000000878

    H E stain (A), in situ hybridization (B) and immunohistochemical staining (C–H) of SARS-CoV2 positive lung (A–E) and SARS-CoV2 negative tissues (F–H); ISH probe to S1 spike protein (B) demonstrating SARS-CoV2 viral RNA in lung tissue(B); SARS-CoV2-positive lung tissues negative for mAb FIPV3-20 (C) and mAb 007 (D); mAb 019, intense immunolabelling of hyaline membranes in lung (E), extensive nonspecific immunolabelling of skin (F), colon (G), and spleen (H).
    Figure Legend Snippet: H E stain (A), in situ hybridization (B) and immunohistochemical staining (C–H) of SARS-CoV2 positive lung (A–E) and SARS-CoV2 negative tissues (F–H); ISH probe to S1 spike protein (B) demonstrating SARS-CoV2 viral RNA in lung tissue(B); SARS-CoV2-positive lung tissues negative for mAb FIPV3-20 (C) and mAb 007 (D); mAb 019, intense immunolabelling of hyaline membranes in lung (E), extensive nonspecific immunolabelling of skin (F), colon (G), and spleen (H).

    Techniques Used: Staining, In Situ Hybridization, Immunohistochemistry

    Pellets of HEK293 cells transfected with SARS-CoV2: S1 and S2 subunit of spike protein, nucleoprotein and untransfected cells. A–D, Hematoxylin eosin stain/H E. E–H, mAb FIPV3-20 to nucleoprotein, no immunolabeling of any pellet. I–L, mAb 019, intense immunostaining of all cell pellets. M–P, mAb 1A9, exclusive immunoreactivity of HEK293 cells transfected with spike protein S2 subunit. Q–T, mAb 001, homogeneous staining of HEK293 cells expressing nucleoprotein. U–X. In situ hybridization with probe to S1 subunit positive in corresponding HEK293 cells.
    Figure Legend Snippet: Pellets of HEK293 cells transfected with SARS-CoV2: S1 and S2 subunit of spike protein, nucleoprotein and untransfected cells. A–D, Hematoxylin eosin stain/H E. E–H, mAb FIPV3-20 to nucleoprotein, no immunolabeling of any pellet. I–L, mAb 019, intense immunostaining of all cell pellets. M–P, mAb 1A9, exclusive immunoreactivity of HEK293 cells transfected with spike protein S2 subunit. Q–T, mAb 001, homogeneous staining of HEK293 cells expressing nucleoprotein. U–X. In situ hybridization with probe to S1 subunit positive in corresponding HEK293 cells.

    Techniques Used: Transfection, Staining, Immunolabeling, Immunostaining, Expressing, In Situ Hybridization

    Analysis of autopsy tissue from patients who died from COVID-19: H E stain (A, D) immunoreactivity with mAb 1A9 to S2 subunit spike protein (B, E) and mAb 001 to nucleoprotein (C, F–I); COVID-19 pneumonia in acute phase diffuse alveolar damage with serial sections stained with H E (A), mAb 1A9 (B), and mAb 001 (C); extensive immunostaining of hyaline membranes and alveolar macrophages for both spike protein and nucleoprotein. mAb 001 staining in endothelia of focal septal vessels and pneumocytes (inset); mucus filled bronchus (D) H E stain with weakly positive S2 spike protein content (E) and strong signal for nucleoprotein (F); heart muscle with mAb 001 immunopositive endothelia; mAb 001/nucleoprotein-positive content in tubule of SARS-CoV2-positive autopsy kidney (H) and smear of nasal swab stained with mAb 001 positive for SARS-CoV2 (I).
    Figure Legend Snippet: Analysis of autopsy tissue from patients who died from COVID-19: H E stain (A, D) immunoreactivity with mAb 1A9 to S2 subunit spike protein (B, E) and mAb 001 to nucleoprotein (C, F–I); COVID-19 pneumonia in acute phase diffuse alveolar damage with serial sections stained with H E (A), mAb 1A9 (B), and mAb 001 (C); extensive immunostaining of hyaline membranes and alveolar macrophages for both spike protein and nucleoprotein. mAb 001 staining in endothelia of focal septal vessels and pneumocytes (inset); mucus filled bronchus (D) H E stain with weakly positive S2 spike protein content (E) and strong signal for nucleoprotein (F); heart muscle with mAb 001 immunopositive endothelia; mAb 001/nucleoprotein-positive content in tubule of SARS-CoV2-positive autopsy kidney (H) and smear of nasal swab stained with mAb 001 positive for SARS-CoV2 (I).

    Techniques Used: Staining, Immunostaining

    16) Product Images from "Identification of Human Single-Domain Antibodies against SARS-CoV-2"

    Article Title: Identification of Human Single-Domain Antibodies against SARS-CoV-2

    Journal: Cell Host & Microbe

    doi: 10.1016/j.chom.2020.04.023

    Characterization of Single-Domain Antibodies Identified from Antibody Library Using SARS-CoV-2 RBD and S1 as Panning Antigens (A) Eighteen single-domain antibodies identified by panning against SARS-CoV-2 RBD and 5 antibodies by using SARS-CoV-2 S1 as panning antigens were tested in competition binding assay. Competition of these antibodies with each other, or ACE2, or the antibody CR3022 for RBD binding were measured by BLI. The antibodies are displayed in 5 groups (A, B, C, D, or E). The values are the percentage of binding that occurred during competition in comparison with non-competed binding, which was normalized to 100%, and the range of competition is indicated by the box colors. Black-filled boxes indicate strongly competing pairs (residual binding
    Figure Legend Snippet: Characterization of Single-Domain Antibodies Identified from Antibody Library Using SARS-CoV-2 RBD and S1 as Panning Antigens (A) Eighteen single-domain antibodies identified by panning against SARS-CoV-2 RBD and 5 antibodies by using SARS-CoV-2 S1 as panning antigens were tested in competition binding assay. Competition of these antibodies with each other, or ACE2, or the antibody CR3022 for RBD binding were measured by BLI. The antibodies are displayed in 5 groups (A, B, C, D, or E). The values are the percentage of binding that occurred during competition in comparison with non-competed binding, which was normalized to 100%, and the range of competition is indicated by the box colors. Black-filled boxes indicate strongly competing pairs (residual binding

    Techniques Used: Binding Assay

    17) Product Images from "CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells"

    Article Title: CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells

    Journal: Signal Transduction and Targeted Therapy

    doi: 10.1038/s41392-020-00426-x

    CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p
    Figure Legend Snippet: CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p

    Techniques Used: Infection, Co-Immunoprecipitation Assay, Microscopy, Immunofluorescence, Multicolor Immunofluorescence Staining, Real-time Polymerase Chain Reaction

    18) Product Images from "Cloning, Expression and Biophysical Characterization of a Yeast-expressed Recombinant SARS-CoV-2 Receptor Binding Domain COVID-19 Vaccine Candidate"

    Article Title: Cloning, Expression and Biophysical Characterization of a Yeast-expressed Recombinant SARS-CoV-2 Receptor Binding Domain COVID-19 Vaccine Candidate

    Journal: bioRxiv

    doi: 10.1101/2020.11.09.373449

    Coomassie Blue stained SDS-PAGE and western blot probed with anti-SARS-CoV-2 Spike rabbit antibody. (A) SDS-PAGE gel of 10 μL fermentation supernatant for tagged-free RBD219-WT, RBD219-N1, and RBD219-N1C1; (B) Coomassie Blue stained SDS-PAGE gel of 3 μg purified RBDs or western blot of 1.5 μg of the purified RBDs under non-reduced and reduced conditions. (C) SDS-PAGE of 3 μg PNGase-F treated purified RBDs; please note that the 37 kDa band observed on the PNGase-F treated gel is the N-glycosidase PNGase-F enzyme.
    Figure Legend Snippet: Coomassie Blue stained SDS-PAGE and western blot probed with anti-SARS-CoV-2 Spike rabbit antibody. (A) SDS-PAGE gel of 10 μL fermentation supernatant for tagged-free RBD219-WT, RBD219-N1, and RBD219-N1C1; (B) Coomassie Blue stained SDS-PAGE gel of 3 μg purified RBDs or western blot of 1.5 μg of the purified RBDs under non-reduced and reduced conditions. (C) SDS-PAGE of 3 μg PNGase-F treated purified RBDs; please note that the 37 kDa band observed on the PNGase-F treated gel is the N-glycosidase PNGase-F enzyme.

    Techniques Used: Staining, SDS Page, Western Blot, Purification

    19) Product Images from "SARS-CoV-2 invades host cells via a novel route: CD147-spike protein"

    Article Title: SARS-CoV-2 invades host cells via a novel route: CD147-spike protein

    Journal: bioRxiv

    doi: 10.1101/2020.03.14.988345

    The localization of CD147 and SP observed by immune-electron microscope. (A) The SARS-CoV-2 infected Vero E6 cell with visible SARS-CoV-2 virion (black arrow). ( B and C ) The localization of CD147 (10nm-gold colloid, yellow arrows) and SP (20nm-gold colloid, red arrows) in viral inclusion bodies of SARS-CoV-2 infected Vero E6 cell. Scale bar = 200nm.
    Figure Legend Snippet: The localization of CD147 and SP observed by immune-electron microscope. (A) The SARS-CoV-2 infected Vero E6 cell with visible SARS-CoV-2 virion (black arrow). ( B and C ) The localization of CD147 (10nm-gold colloid, yellow arrows) and SP (20nm-gold colloid, red arrows) in viral inclusion bodies of SARS-CoV-2 infected Vero E6 cell. Scale bar = 200nm.

    Techniques Used: Microscopy, Infection

    Meplazumab inhibits the SARS-CoV-2 replication. (A and B) 1×10 4 Vero E6 cells were cultured in a 96-well plate at 37°C overnight, the supernatant was discard and 100 μl of medium (containing different concentrations of Meplazumab) was added into the plates to incubate for 1 h. Then the cells were infected with SARS-CoV-2 (100TCID 50 ). After one-hour infection, the supernatants were removed and 200 μl of medium (containing different concentrations of Meplazumab) was added, the cells were cultured to observe the cytopathic changes for 2-3 days. The supernatants were harvested to detect the gene copy number of virus with quantitative PCR and the Vero E6 cells were stained by crystal violet staining. Finally, the value of optical density (OD) at 570 nm was measured with a microplate reader.
    Figure Legend Snippet: Meplazumab inhibits the SARS-CoV-2 replication. (A and B) 1×10 4 Vero E6 cells were cultured in a 96-well plate at 37°C overnight, the supernatant was discard and 100 μl of medium (containing different concentrations of Meplazumab) was added into the plates to incubate for 1 h. Then the cells were infected with SARS-CoV-2 (100TCID 50 ). After one-hour infection, the supernatants were removed and 200 μl of medium (containing different concentrations of Meplazumab) was added, the cells were cultured to observe the cytopathic changes for 2-3 days. The supernatants were harvested to detect the gene copy number of virus with quantitative PCR and the Vero E6 cells were stained by crystal violet staining. Finally, the value of optical density (OD) at 570 nm was measured with a microplate reader.

    Techniques Used: Cell Culture, Infection, Real-time Polymerase Chain Reaction, Staining

    Identification of interaction between CD147 and SP. (A) The interaction of CD147 and SP detected by SPR assay, KD = 1.85×10 −7 M. (B) The interaction of CD147 and SP detected by Co-IP assay. Anti-CD147 antibody and anti-SARS-CoV-2 Spike antibody were used for antibody immobilization for Co-IP. The mIgG and rIgG were selected as negative control. (C) The interaction of CD147 and SP detected by ELISA. (D) The ability of Meplazumab to compete with SP for CD147 binding performed by competitive inhibition ELISA.
    Figure Legend Snippet: Identification of interaction between CD147 and SP. (A) The interaction of CD147 and SP detected by SPR assay, KD = 1.85×10 −7 M. (B) The interaction of CD147 and SP detected by Co-IP assay. Anti-CD147 antibody and anti-SARS-CoV-2 Spike antibody were used for antibody immobilization for Co-IP. The mIgG and rIgG were selected as negative control. (C) The interaction of CD147 and SP detected by ELISA. (D) The ability of Meplazumab to compete with SP for CD147 binding performed by competitive inhibition ELISA.

    Techniques Used: SPR Assay, Co-Immunoprecipitation Assay, Negative Control, Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition

    20) Product Images from "Rapid Detection of COVID-19 Causative Virus (SARS-CoV-2) in Human Nasopharyngeal Swab Specimens Using Field-Effect Transistor-Based Biosensor"

    Article Title: Rapid Detection of COVID-19 Causative Virus (SARS-CoV-2) in Human Nasopharyngeal Swab Specimens Using Field-Effect Transistor-Based Biosensor

    Journal: ACS Nano

    doi: 10.1021/acsnano.0c02823

    Detection of SARS-CoV-2 antigen protein. (A) Schematic diagram for the COVID-19 FET sensor for detection of SARS-CoV-2 spike protein. (B) Real-time response of COVID-19 FET toward SARS-CoV-2 antigen protein in PBS and (C) related dose-dependent response curve ( V DS = 0.01 V). Graphene-based FET without SARS-CoV-2 antibody is presented as negative control. (D) Selective response of COVID-19 FET sensor toward target SARS-CoV-2 antigen protein and MERS-CoV protein. (E) Real-time response of COVID-19 FET toward SARS-CoV-2 antigen protein in UTM and (F) related dose-dependent response curve.
    Figure Legend Snippet: Detection of SARS-CoV-2 antigen protein. (A) Schematic diagram for the COVID-19 FET sensor for detection of SARS-CoV-2 spike protein. (B) Real-time response of COVID-19 FET toward SARS-CoV-2 antigen protein in PBS and (C) related dose-dependent response curve ( V DS = 0.01 V). Graphene-based FET without SARS-CoV-2 antibody is presented as negative control. (D) Selective response of COVID-19 FET sensor toward target SARS-CoV-2 antigen protein and MERS-CoV protein. (E) Real-time response of COVID-19 FET toward SARS-CoV-2 antigen protein in UTM and (F) related dose-dependent response curve.

    Techniques Used: Negative Control

    Detection of cultured SARS-CoV-2 virus. (A) Schematic diagram for the COVID-19 FET sensor for detection of SARS-CoV-2 cultured virus. (B) Real-time response of COVID-19 FET toward SARS-CoV-2 cultured virus and (C) related dose-dependent response curve.
    Figure Legend Snippet: Detection of cultured SARS-CoV-2 virus. (A) Schematic diagram for the COVID-19 FET sensor for detection of SARS-CoV-2 cultured virus. (B) Real-time response of COVID-19 FET toward SARS-CoV-2 cultured virus and (C) related dose-dependent response curve.

    Techniques Used: Cell Culture

    Electrical characterization of pristine, PBASE-modified, and SARS-CoV-2 spike antibody-immobilized graphene. (A) Schematic diagram of the aqueous-solution-gated FET (COVID-19 FET sensor) configuration using the antibody-conjugated graphene. (B) I DS – V DS output curves of the antibody-conjugated FET with various gating voltages from 0 to −1.5 V in steps of −0.3 V. I DS negatively increased as V GS negatively increased. (C) Current–voltage ( I–V ) characteristics of the graphene-based device of each functionalization process for the antibody modification. (D) Measurement of transfer curves of the COVID-19 FET sensor in steps of the antibody conjugation ( V DS = 0.01 V).
    Figure Legend Snippet: Electrical characterization of pristine, PBASE-modified, and SARS-CoV-2 spike antibody-immobilized graphene. (A) Schematic diagram of the aqueous-solution-gated FET (COVID-19 FET sensor) configuration using the antibody-conjugated graphene. (B) I DS – V DS output curves of the antibody-conjugated FET with various gating voltages from 0 to −1.5 V in steps of −0.3 V. I DS negatively increased as V GS negatively increased. (C) Current–voltage ( I–V ) characteristics of the graphene-based device of each functionalization process for the antibody modification. (D) Measurement of transfer curves of the COVID-19 FET sensor in steps of the antibody conjugation ( V DS = 0.01 V).

    Techniques Used: Modification, Conjugation Assay

    Detection of SARS-CoV-2 virus from clinical samples. (A) Schematic diagram for the COVID-19 FET sensor for detection of SARS-CoV-2 virus from COVID-19 patients. (B,C) Comparison of response signal between normal samples and patient ones. (D) Real-time response of COVID-19 FET toward SARS-CoV-2 clinical sample and (C) related dose-dependent response curve.
    Figure Legend Snippet: Detection of SARS-CoV-2 virus from clinical samples. (A) Schematic diagram for the COVID-19 FET sensor for detection of SARS-CoV-2 virus from COVID-19 patients. (B,C) Comparison of response signal between normal samples and patient ones. (D) Real-time response of COVID-19 FET toward SARS-CoV-2 clinical sample and (C) related dose-dependent response curve.

    Techniques Used:

    21) Product Images from "Rapid and quantitative detection of SARS-CoV-2 specific IgG for convalescent serum evaluation"

    Article Title: Rapid and quantitative detection of SARS-CoV-2 specific IgG for convalescent serum evaluation

    Journal: Biosensors & Bioelectronics

    doi: 10.1016/j.bios.2020.112572

    Affinity screening of the calibration antibodies. (A) Calibration curves of 4 different monoclonal humanized S1 specific IgG against the S1 protein from SARS-CoV-2. (B) Calibration curves of 4 different monoclonal humanized S1 specific IgG against the S1 protein from SARS-CoV (B). The solid lines are the linear fit of the data in the log-log scale. D006 is the only antibody that has a high affinity and high specificity towards SARS-CoV-2 S1. Illustration of the assay mechanism, which uses a single-step ELISA format, is shown in Fig. 1 (A). The sample-to-answer time of this assay is 8 min.
    Figure Legend Snippet: Affinity screening of the calibration antibodies. (A) Calibration curves of 4 different monoclonal humanized S1 specific IgG against the S1 protein from SARS-CoV-2. (B) Calibration curves of 4 different monoclonal humanized S1 specific IgG against the S1 protein from SARS-CoV (B). The solid lines are the linear fit of the data in the log-log scale. D006 is the only antibody that has a high affinity and high specificity towards SARS-CoV-2 S1. Illustration of the assay mechanism, which uses a single-step ELISA format, is shown in Fig. 1 (A). The sample-to-answer time of this assay is 8 min.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    SARS-CoV-2 antigen detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 40 min. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3 × standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein is 0.004 ng/mL
    Figure Legend Snippet: SARS-CoV-2 antigen detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 40 min. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3 × standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein is 0.004 ng/mL

    Techniques Used: Standard Deviation

    Graphical illustrations of the COVID-19 related immunoassays that were performed with our microfluidic chemiluminescent ELISA platform, including (A) affinity evaluation of calibration antibodies, (B) detection of circulating anti-SARS-CoV-2 S1 IgG in serum samples, and (C) detection of SARS-CoV-2 antigens such as S1 and N protein.
    Figure Legend Snippet: Graphical illustrations of the COVID-19 related immunoassays that were performed with our microfluidic chemiluminescent ELISA platform, including (A) affinity evaluation of calibration antibodies, (B) detection of circulating anti-SARS-CoV-2 S1 IgG in serum samples, and (C) detection of SARS-CoV-2 antigens such as S1 and N protein.

    Techniques Used: Chemiluminescent ELISA

    22) Product Images from "Cloning, Expression and Biophysical Characterization of a Yeast-expressed Recombinant SARS-CoV-2 Receptor Binding Domain COVID-19 Vaccine Candidate"

    Article Title: Cloning, Expression and Biophysical Characterization of a Yeast-expressed Recombinant SARS-CoV-2 Receptor Binding Domain COVID-19 Vaccine Candidate

    Journal: bioRxiv

    doi: 10.1101/2020.11.09.373449

    Coomassie Blue stained SDS-PAGE and western blot probed with anti-SARS-CoV-2 Spike rabbit antibody. (A) SDS-PAGE gel of 10 μL fermentation supernatant for tagged-free RBD219-WT, RBD219-N1, and RBD219-N1C1; (B) Coomassie Blue stained SDS-PAGE gel of 3 μg purified RBDs or western blot of 1.5 μg of the purified RBDs under non-reduced and reduced conditions. (C) SDS-PAGE of 3 μg PNGase-F treated purified RBDs; please note that the 37 kDa band observed on the PNGase-F treated gel is the N-glycosidase PNGase-F enzyme.
    Figure Legend Snippet: Coomassie Blue stained SDS-PAGE and western blot probed with anti-SARS-CoV-2 Spike rabbit antibody. (A) SDS-PAGE gel of 10 μL fermentation supernatant for tagged-free RBD219-WT, RBD219-N1, and RBD219-N1C1; (B) Coomassie Blue stained SDS-PAGE gel of 3 μg purified RBDs or western blot of 1.5 μg of the purified RBDs under non-reduced and reduced conditions. (C) SDS-PAGE of 3 μg PNGase-F treated purified RBDs; please note that the 37 kDa band observed on the PNGase-F treated gel is the N-glycosidase PNGase-F enzyme.

    Techniques Used: Staining, SDS Page, Western Blot, Purification

    23) Product Images from "Immune responses to SARS-CoV-2 in three children of parents with symptomatic COVID-19"

    Article Title: Immune responses to SARS-CoV-2 in three children of parents with symptomatic COVID-19

    Journal: Nature Communications

    doi: 10.1038/s41467-020-19545-8

    Salivary and plasma antibody responses against SARS-CoV-2 S1 protein by ELISA and by microneutralization assay. a Anti-S1 salivary IgA, IgG, and IgM. # IgA anti-S1 response that developed concurrent with resolution of symptoms. b Anti-S1 plasma IgA, IgG, and IgM. c Neutralizing antibody activity in plasma. A1: mother, A2: father, C1: male (9 years), C2: male (7 years), C3: female (5 years), (P) positive control.
    Figure Legend Snippet: Salivary and plasma antibody responses against SARS-CoV-2 S1 protein by ELISA and by microneutralization assay. a Anti-S1 salivary IgA, IgG, and IgM. # IgA anti-S1 response that developed concurrent with resolution of symptoms. b Anti-S1 plasma IgA, IgG, and IgM. c Neutralizing antibody activity in plasma. A1: mother, A2: father, C1: male (9 years), C2: male (7 years), C3: female (5 years), (P) positive control.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Microneutralization Assay, Activity Assay, Positive Control

    24) Product Images from "Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients"

    Article Title: Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients

    Journal: Biosensors & Bioelectronics

    doi: 10.1016/j.bios.2020.112643

    Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) Array exposed 20% FBS + 10% PNHS; (B) Array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Ångstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.
    Figure Legend Snippet: Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) Array exposed 20% FBS + 10% PNHS; (B) Array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Ångstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.

    Techniques Used: Titration, Concentration Assay, Standard Deviation

    25) Product Images from "Identification of Immunohistochemical Reagents for In Situ Protein Expression Analysis of Coronavirus-associated Changes in Human Tissues"

    Article Title: Identification of Immunohistochemical Reagents for In Situ Protein Expression Analysis of Coronavirus-associated Changes in Human Tissues

    Journal: Applied Immunohistochemistry & Molecular Morphology

    doi: 10.1097/PAI.0000000000000878

    Pellets of HEK293 cells transfected with SARS-CoV2: S1 and S2 subunit of spike protein, nucleoprotein and untransfected cells. A–D, Hematoxylin eosin stain/H  E. E–H, mAb FIPV3-20 to nucleoprotein, no immunolabeling of any pellet. I–L, mAb 019, intense immunostaining of all cell pellets. M–P, mAb 1A9, exclusive immunoreactivity of HEK293 cells transfected with spike protein S2 subunit. Q–T, mAb 001, homogeneous staining of HEK293 cells expressing nucleoprotein. U–X. In situ hybridization with probe to S1 subunit positive in corresponding HEK293 cells.
    Figure Legend Snippet: Pellets of HEK293 cells transfected with SARS-CoV2: S1 and S2 subunit of spike protein, nucleoprotein and untransfected cells. A–D, Hematoxylin eosin stain/H E. E–H, mAb FIPV3-20 to nucleoprotein, no immunolabeling of any pellet. I–L, mAb 019, intense immunostaining of all cell pellets. M–P, mAb 1A9, exclusive immunoreactivity of HEK293 cells transfected with spike protein S2 subunit. Q–T, mAb 001, homogeneous staining of HEK293 cells expressing nucleoprotein. U–X. In situ hybridization with probe to S1 subunit positive in corresponding HEK293 cells.

    Techniques Used: Transfection, Staining, Immunolabeling, Immunostaining, Expressing, In Situ Hybridization

    26) Product Images from "An all-solid-state heterojunction oxide transistor for the rapid detection of biomolecules and SARS-CoV-2 spike S1 protein"

    Article Title: An all-solid-state heterojunction oxide transistor for the rapid detection of biomolecules and SARS-CoV-2 spike S1 protein

    Journal: bioRxiv

    doi: 10.1101/2021.01.19.427256

    Detection of SARS-CoV-2 spike protein. a , Schematic of the SARS-CoV-2 spike S1 protein detection. The SARS-CoV-2 spike S1 antibody is anchored onto the sensor platform after the sequential modification of oxide surface with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde. b , Transfer characteristics (V D = 3 V) of a fully functionalised tri-channel transistor sensor measured in the presence of the SARS-CoV-2 spike protein in 0.1× phosphate-buffered saline (PBS, baseline). c , Real-time response of the tri-channel transistor sensors to different concentrations (1 fM to 100 pM) of the SARS-CoV-2 spike protein and the MERS-CoV protein in 0.1× PBS.
    Figure Legend Snippet: Detection of SARS-CoV-2 spike protein. a , Schematic of the SARS-CoV-2 spike S1 protein detection. The SARS-CoV-2 spike S1 antibody is anchored onto the sensor platform after the sequential modification of oxide surface with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde. b , Transfer characteristics (V D = 3 V) of a fully functionalised tri-channel transistor sensor measured in the presence of the SARS-CoV-2 spike protein in 0.1× phosphate-buffered saline (PBS, baseline). c , Real-time response of the tri-channel transistor sensors to different concentrations (1 fM to 100 pM) of the SARS-CoV-2 spike protein and the MERS-CoV protein in 0.1× PBS.

    Techniques Used: Modification

    27) Product Images from "An all-solid-state heterojunction oxide transistor for the rapid detection of biomolecules and SARS-CoV-2 spike S1 protein"

    Article Title: An all-solid-state heterojunction oxide transistor for the rapid detection of biomolecules and SARS-CoV-2 spike S1 protein

    Journal: bioRxiv

    doi: 10.1101/2021.01.19.427256

    Detection of SARS-CoV-2 spike protein. a , Schematic of the SARS-CoV-2 spike S1 protein detection. The SARS-CoV-2 spike S1 antibody is anchored onto the sensor platform after the sequential modification of oxide surface with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde. b , Transfer characteristics (V D = 3 V) of a fully functionalised tri-channel transistor sensor measured in the presence of the SARS-CoV-2 spike protein in 0.1× phosphate-buffered saline (PBS, baseline). c , Real-time response of the tri-channel transistor sensors to different concentrations (1 fM to 100 pM) of the SARS-CoV-2 spike protein and the MERS-CoV protein in 0.1× PBS.
    Figure Legend Snippet: Detection of SARS-CoV-2 spike protein. a , Schematic of the SARS-CoV-2 spike S1 protein detection. The SARS-CoV-2 spike S1 antibody is anchored onto the sensor platform after the sequential modification of oxide surface with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde. b , Transfer characteristics (V D = 3 V) of a fully functionalised tri-channel transistor sensor measured in the presence of the SARS-CoV-2 spike protein in 0.1× phosphate-buffered saline (PBS, baseline). c , Real-time response of the tri-channel transistor sensors to different concentrations (1 fM to 100 pM) of the SARS-CoV-2 spike protein and the MERS-CoV protein in 0.1× PBS.

    Techniques Used: Modification

    28) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    29) Product Images from "Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples"

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    Journal: bioRxiv

    doi: 10.1101/2020.04.20.052233

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Figure Legend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Techniques Used: Generated

    30) Product Images from "Rapid and sensitive detection of SARS-CoV-2 antibodies by biolayer interferometry"

    Article Title: Rapid and sensitive detection of SARS-CoV-2 antibodies by biolayer interferometry

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-78895-x

    BLI-ISA evaluation of SARS-CoV-2 spike RBD reactivity of pre-pandemic and convalescent plasma. ( a , b ) Single-dilution BLI-ISA to evaluate the presence of RBD-reactive human antibodies in the pre-pandemic seronegative (SN, cyan) and convalescent seropositive (SP, red) samples compared to no-antigen controls (grey). The assays were performed with plasma at a 1:8 dilution. Bars and dots represent the mean of biological duplicates, and error bars represent one standard deviation from the mean. Blue and green dashed lines represent the mean of seronegative samples plus 3 and 5 standard deviations, respectively. ( a ) The Total Antibody Binding signal is measured when RBD-biotin-loaded SA biosensors are dipped into plasma samples. ( b ) The Detection signal is measured when RBD-biotin-loaded SA biosensors that had been dipped into plasma are subsequently dipped into colloidal gold-conjugated anti-human IgG. ( c ) Dilution series BLI-ISA from representative strong (SP7) and moderate (SP8) seropositive samples. ( d ) Dilution series BLI-ISA from the weakest seropositive sample (SP3) compared to seronegative plasma samples.
    Figure Legend Snippet: BLI-ISA evaluation of SARS-CoV-2 spike RBD reactivity of pre-pandemic and convalescent plasma. ( a , b ) Single-dilution BLI-ISA to evaluate the presence of RBD-reactive human antibodies in the pre-pandemic seronegative (SN, cyan) and convalescent seropositive (SP, red) samples compared to no-antigen controls (grey). The assays were performed with plasma at a 1:8 dilution. Bars and dots represent the mean of biological duplicates, and error bars represent one standard deviation from the mean. Blue and green dashed lines represent the mean of seronegative samples plus 3 and 5 standard deviations, respectively. ( a ) The Total Antibody Binding signal is measured when RBD-biotin-loaded SA biosensors are dipped into plasma samples. ( b ) The Detection signal is measured when RBD-biotin-loaded SA biosensors that had been dipped into plasma are subsequently dipped into colloidal gold-conjugated anti-human IgG. ( c ) Dilution series BLI-ISA from representative strong (SP7) and moderate (SP8) seropositive samples. ( d ) Dilution series BLI-ISA from the weakest seropositive sample (SP3) compared to seronegative plasma samples.

    Techniques Used: Standard Deviation, Binding Assay

    ELISA evaluation of SARS-CoV-2 spike RBD reactivity of pre-pandemic and convalescent plasma. ( a ) Single-dilution ELISA to evaluate the presence of RBD-reactive human IgG in pre-pandemic seronegative (SN, cyan) and convalescent seropositive (SP, red) samples compared to no-antigen controls (grey). The assays were performed with plasma at a 1:50 dilution. Samples were evaluated in biological duplicates and error bars represent one standard deviation from the mean. Blue and green dashed lines represent the mean of seronegative samples plus 3 and 5 standard deviations, respectively. ( b ) Dilution series ELISA was performed to quantitate RBD-reactive human IgG in plasma. Samples were evaluated in biological duplicates. Dashed curves represent fit lines from a four-parameter logistic regression applied over each series. ( c ) Data from ( b ) plotted as area-under-the-curve (AUC).
    Figure Legend Snippet: ELISA evaluation of SARS-CoV-2 spike RBD reactivity of pre-pandemic and convalescent plasma. ( a ) Single-dilution ELISA to evaluate the presence of RBD-reactive human IgG in pre-pandemic seronegative (SN, cyan) and convalescent seropositive (SP, red) samples compared to no-antigen controls (grey). The assays were performed with plasma at a 1:50 dilution. Samples were evaluated in biological duplicates and error bars represent one standard deviation from the mean. Blue and green dashed lines represent the mean of seronegative samples plus 3 and 5 standard deviations, respectively. ( b ) Dilution series ELISA was performed to quantitate RBD-reactive human IgG in plasma. Samples were evaluated in biological duplicates. Dashed curves represent fit lines from a four-parameter logistic regression applied over each series. ( c ) Data from ( b ) plotted as area-under-the-curve (AUC).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Standard Deviation

    BLI-ISA evaluation of plasma antibodies to SARS-CoV-2 prefusion Spike and plasma IgA to SARS-CoV-2 spike RBD. ( a ) Single-dilution BLI-ISA to evaluate the presence of prefusion Spike-reactive human antibodies in the pre-pandemic seronegative (SN, cyan) and convalescent seropositive (SP, red) samples. The Total Antibody Binding signal (left) is measured when prefusion Spike-His-loaded HIS1K biosensors are dipped into plasma samples. The Detection signal (right) is measured when prefusion Spike-His-loaded HIS1K biosensors that had been dipped into plasma are subsequently dipped into colloidal gold-conjugated anti-human IgG. ( b ) Single-dilution BLI-ISA to evaluate the presence of RBD-reactive human antibodies in the samples. The Total Antibody Binding signal (left) is measured when RBD-biotin-loaded SA biosensors are dipped into plasma samples. The Detection signal (right) is measured when RBD-biotin-loaded SA biosensors that had been dipped into plasma are subsequently dipped into colloidal gold-conjugated anti-human IgA. The SP7 dot is colored pink to indicate that this sample had a negative signal (value in parentheses) in the Detection step. All assays were performed with plasma at a 1:8 dilution. Dots represent the mean of biological duplicates, and error bars represent one standard deviation from the mean.
    Figure Legend Snippet: BLI-ISA evaluation of plasma antibodies to SARS-CoV-2 prefusion Spike and plasma IgA to SARS-CoV-2 spike RBD. ( a ) Single-dilution BLI-ISA to evaluate the presence of prefusion Spike-reactive human antibodies in the pre-pandemic seronegative (SN, cyan) and convalescent seropositive (SP, red) samples. The Total Antibody Binding signal (left) is measured when prefusion Spike-His-loaded HIS1K biosensors are dipped into plasma samples. The Detection signal (right) is measured when prefusion Spike-His-loaded HIS1K biosensors that had been dipped into plasma are subsequently dipped into colloidal gold-conjugated anti-human IgG. ( b ) Single-dilution BLI-ISA to evaluate the presence of RBD-reactive human antibodies in the samples. The Total Antibody Binding signal (left) is measured when RBD-biotin-loaded SA biosensors are dipped into plasma samples. The Detection signal (right) is measured when RBD-biotin-loaded SA biosensors that had been dipped into plasma are subsequently dipped into colloidal gold-conjugated anti-human IgA. The SP7 dot is colored pink to indicate that this sample had a negative signal (value in parentheses) in the Detection step. All assays were performed with plasma at a 1:8 dilution. Dots represent the mean of biological duplicates, and error bars represent one standard deviation from the mean.

    Techniques Used: Binding Assay, Standard Deviation

    31) Product Images from "Prunella vulgaris extract and suramin block SARS-coronavirus 2 virus Spike protein D614 and G614 variants mediated receptor association and virus entry in cell culture system"

    Article Title: Prunella vulgaris extract and suramin block SARS-coronavirus 2 virus Spike protein D614 and G614 variants mediated receptor association and virus entry in cell culture system

    Journal: bioRxiv

    doi: 10.1101/2020.08.28.270306

    SARS-CoV-2 SP-PVs’s infection in different cell lines and SARS-CoV-2 SP G614 variant exhibited stronger virus entry. A) 293T, 293T ACE2 and Vero-E6 cells were infected by equal amounts of SARS-CoV-2SP-, SARS-CoV-2SPΔC-pseudotyped viruses. At 48 hrs pi, the Gluc activity in supernatants was measured. B) the expression of SARS-CoV-2SP receptor, ACE2, in 293T, 293T ACE2 and Vero-E6 cells detected by WB with anti-ACE2 antibodies. C) The SPΔC G614 -GFP + PVs were produced with 293T cells and used to infect 293T ACE2 cells in 96-well plate After 48 hrs pi, GFP-positive cells (per well) were counted and photographed by fluorescence microscope (on the top of the panel). D) Detection of SARS-CoV-2 SPΔC, SPΔC G614 and HIV p24 protein expression in transfected 293T cells and viral particles by WB. E) Infectivity comparison of SPΔC-PVs and SPΔC G614 -PVs in 293T ACE2 cells. Equal amounts of SPΔC D614 -PVs and SPΔC G614 -PVs virions (adjusted by p24 level) were used to infect 293T ACE2 cells. At different days post-infection (pi), Gluc activity in supernatants was measured.
    Figure Legend Snippet: SARS-CoV-2 SP-PVs’s infection in different cell lines and SARS-CoV-2 SP G614 variant exhibited stronger virus entry. A) 293T, 293T ACE2 and Vero-E6 cells were infected by equal amounts of SARS-CoV-2SP-, SARS-CoV-2SPΔC-pseudotyped viruses. At 48 hrs pi, the Gluc activity in supernatants was measured. B) the expression of SARS-CoV-2SP receptor, ACE2, in 293T, 293T ACE2 and Vero-E6 cells detected by WB with anti-ACE2 antibodies. C) The SPΔC G614 -GFP + PVs were produced with 293T cells and used to infect 293T ACE2 cells in 96-well plate After 48 hrs pi, GFP-positive cells (per well) were counted and photographed by fluorescence microscope (on the top of the panel). D) Detection of SARS-CoV-2 SPΔC, SPΔC G614 and HIV p24 protein expression in transfected 293T cells and viral particles by WB. E) Infectivity comparison of SPΔC-PVs and SPΔC G614 -PVs in 293T ACE2 cells. Equal amounts of SPΔC D614 -PVs and SPΔC G614 -PVs virions (adjusted by p24 level) were used to infect 293T ACE2 cells. At different days post-infection (pi), Gluc activity in supernatants was measured.

    Techniques Used: Infection, Variant Assay, Activity Assay, Expressing, Western Blot, Produced, Fluorescence, Microscopy, Transfection

    SARS-CoV-2-SP-PV’s infection was efficiently blocked by CHPV and suramin. A) Images of the dried Prunella Vulgaris flowers and its water extract (CHPV). B) Dose -response anti-SARS-CoV-2 analysis by Gluc activity for CHPV or suramin. 293T ACE2 cells were infected by equal amounts of SARS-CoV-2SPΔC-pseudotyped viruses in the presence of different dose of CHPV or suramin. At 48 hrs pi, the Gluc activity in supernatants was measured. (% inhibition = 100 ⨯ [1 - (Gluc value in presence of drug)/(Gluc value in absence of drug)). C) Infection inhibition of CHPV or suramin on SARS-CoV-2-SPΔC G614 -PVs in 293T ACE2 cells. Equal amounts of SCoV-2-SPΔC G614 -PVs (adjusted by p24 level) were used to infect 293T ACE2 cells in presence of different concentrations of CHPV or suramin, in indicated at bottom of the panel. At 48 hrs pi, Gluc activity in supernatants was measured and present as % inhibition. Means ± SD were calculated from duplicate experiments. D) 293T ACE2 cells in 96-well plate were infected with SPΔC G614 -GFP + PVs. After 48 hrs pi, GFP-positive cells (per well) were counted (left panel) and photographed by fluorescence microscope (right panel, a. Without drugs; b. Without infection; c. In the presence of CHPV (100 μg/ml ) ; d. In the presence of suramin (100 μg/ml ) .
    Figure Legend Snippet: SARS-CoV-2-SP-PV’s infection was efficiently blocked by CHPV and suramin. A) Images of the dried Prunella Vulgaris flowers and its water extract (CHPV). B) Dose -response anti-SARS-CoV-2 analysis by Gluc activity for CHPV or suramin. 293T ACE2 cells were infected by equal amounts of SARS-CoV-2SPΔC-pseudotyped viruses in the presence of different dose of CHPV or suramin. At 48 hrs pi, the Gluc activity in supernatants was measured. (% inhibition = 100 ⨯ [1 - (Gluc value in presence of drug)/(Gluc value in absence of drug)). C) Infection inhibition of CHPV or suramin on SARS-CoV-2-SPΔC G614 -PVs in 293T ACE2 cells. Equal amounts of SCoV-2-SPΔC G614 -PVs (adjusted by p24 level) were used to infect 293T ACE2 cells in presence of different concentrations of CHPV or suramin, in indicated at bottom of the panel. At 48 hrs pi, Gluc activity in supernatants was measured and present as % inhibition. Means ± SD were calculated from duplicate experiments. D) 293T ACE2 cells in 96-well plate were infected with SPΔC G614 -GFP + PVs. After 48 hrs pi, GFP-positive cells (per well) were counted (left panel) and photographed by fluorescence microscope (right panel, a. Without drugs; b. Without infection; c. In the presence of CHPV (100 μg/ml ) ; d. In the presence of suramin (100 μg/ml ) .

    Techniques Used: Infection, Activity Assay, Inhibition, Fluorescence, Microscopy

    Characterization of the mechanisms of CHPV and suramin for their anti-SARS-COV-2-SP action. A) Time-dependent inhibition of SPΔC G614 -PVs infection mediated by CHPV or suramin. CHPV (100 μg/mL) or suramin (100 μg/mL) was added at 1 hr prior to infection, during infection (0 hr), and at 1 hr, and 3 hr pi. The positive controls (PC) were 293T ACE2 cells infected with SPΔC G614 -PVs in the absence of compounds. At 3 hrs pi, all of the cell cultures were replaced with fresh DMEM and cultured for 48 hrs. Then, the Gluc activity was monitored in the supernatant, and the data are shown as a percentage of inhibition (%). B) inhibitory effect of CHPV or suramin on SARS-CoV2-SP/ACE2 binding by ELISA as described in materials and methods. nAB: anti-COVID-19 neutralizing antibody (SAD-S35). The results are the mean ± SD of duplicate samples, and the data are representative of results obtained in two independent experiments.
    Figure Legend Snippet: Characterization of the mechanisms of CHPV and suramin for their anti-SARS-COV-2-SP action. A) Time-dependent inhibition of SPΔC G614 -PVs infection mediated by CHPV or suramin. CHPV (100 μg/mL) or suramin (100 μg/mL) was added at 1 hr prior to infection, during infection (0 hr), and at 1 hr, and 3 hr pi. The positive controls (PC) were 293T ACE2 cells infected with SPΔC G614 -PVs in the absence of compounds. At 3 hrs pi, all of the cell cultures were replaced with fresh DMEM and cultured for 48 hrs. Then, the Gluc activity was monitored in the supernatant, and the data are shown as a percentage of inhibition (%). B) inhibitory effect of CHPV or suramin on SARS-CoV2-SP/ACE2 binding by ELISA as described in materials and methods. nAB: anti-COVID-19 neutralizing antibody (SAD-S35). The results are the mean ± SD of duplicate samples, and the data are representative of results obtained in two independent experiments.

    Techniques Used: Inhibition, Infection, Cell Culture, Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay

    Generation of a SARS-COV2-SP-pseudotyped lentiviruse particles (SCoV-2-SP-PVs). A) Schematic representation of SARS-CoV-2SP, SARS-CoV-2SPΔC, and SARS-CoV-2SP G614 ΔC expressing plasmids. B) Schematic representation of plasmids and and procedures for production of SARS-COV2-SP-pseudotyped lentivirus particles (SCoV-2-SP-PVs). C) Detection of SARS-CoV-2 SPs and HIV p24 protein expression in transfected 293T cells and viral particles by Western blot (WB) with anti-SP or anti-p24 antibodies. D) Different amounts of SCoV-2-SP-PVs and SCoV-2-SPΔC-PVs virions (adjusted by p24) were used to infect 293T ACE2 cells. At different time intervels, the Gaussia Luciferase activity (Gluc) (left panel) and PVs-associated p24 (at 72 hrs) in supernatants was measured.
    Figure Legend Snippet: Generation of a SARS-COV2-SP-pseudotyped lentiviruse particles (SCoV-2-SP-PVs). A) Schematic representation of SARS-CoV-2SP, SARS-CoV-2SPΔC, and SARS-CoV-2SP G614 ΔC expressing plasmids. B) Schematic representation of plasmids and and procedures for production of SARS-COV2-SP-pseudotyped lentivirus particles (SCoV-2-SP-PVs). C) Detection of SARS-CoV-2 SPs and HIV p24 protein expression in transfected 293T cells and viral particles by Western blot (WB) with anti-SP or anti-p24 antibodies. D) Different amounts of SCoV-2-SP-PVs and SCoV-2-SPΔC-PVs virions (adjusted by p24) were used to infect 293T ACE2 cells. At different time intervels, the Gaussia Luciferase activity (Gluc) (left panel) and PVs-associated p24 (at 72 hrs) in supernatants was measured.

    Techniques Used: Expressing, Transfection, Western Blot, Luciferase, Activity Assay

    Inhibitory effect of CHPV and Suramin on SARS-CoV-2 infection-induced cytopathic effects. Vero cells were infected with a wild type SARS-CoV-2 virus (hCoV-19/Canada/ON-VIDO-01/2020) in the presence or absence of different concentrations of CHPV and Suramin. After 72 hrs pi., the SARS-CoV-2 infection-induced cytopathic effects in Vero cells were monitored. Error bars represent variation between triplicate samples, and the data of (A) and (B) are representative of results obtained in two independent experiments.
    Figure Legend Snippet: Inhibitory effect of CHPV and Suramin on SARS-CoV-2 infection-induced cytopathic effects. Vero cells were infected with a wild type SARS-CoV-2 virus (hCoV-19/Canada/ON-VIDO-01/2020) in the presence or absence of different concentrations of CHPV and Suramin. After 72 hrs pi., the SARS-CoV-2 infection-induced cytopathic effects in Vero cells were monitored. Error bars represent variation between triplicate samples, and the data of (A) and (B) are representative of results obtained in two independent experiments.

    Techniques Used: Infection

    32) Product Images from "CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells"

    Article Title: CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells

    Journal: Signal Transduction and Targeted Therapy

    doi: 10.1038/s41392-020-00426-x

    CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p
    Figure Legend Snippet: CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p

    Techniques Used: Infection, Co-Immunoprecipitation Assay, Microscopy, Immunofluorescence, Multicolor Immunofluorescence Staining, Real-time Polymerase Chain Reaction

    SARS-CoV-2 enters the host cells through CD147-mediated endocytosis. a The sequential endocytosis of SARS-CoV-2 was observed in Vero E6 cells by electron microscope. Scale bars: 200 nm. b The co-localization of spike protein, CD147, and Rab5 were analyzed in BHK-21-CD147 cells and lung tissues from COVID-19 patient by multicolor immunofluorescence staining. Magnification: ×200.
    Figure Legend Snippet: SARS-CoV-2 enters the host cells through CD147-mediated endocytosis. a The sequential endocytosis of SARS-CoV-2 was observed in Vero E6 cells by electron microscope. Scale bars: 200 nm. b The co-localization of spike protein, CD147, and Rab5 were analyzed in BHK-21-CD147 cells and lung tissues from COVID-19 patient by multicolor immunofluorescence staining. Magnification: ×200.

    Techniques Used: Microscopy, Multicolor Immunofluorescence Staining

    Identification of the interaction and co-localization between CD147 and spike protein. a – c The interaction of CD147 and spike was detected by SPR assay ( a ), ELISA ( b ), and Co-IP assay ( c ). The mouse IgG (mIgG) and rabbit IgG (rIgG) were served as negative controls. d OpNS-EM images of CD147, spike(RBD) and CD147-spike(RBD) complexes. Left panels showed the survey view of the micrograph. Right panels showed 8 class averages were selected from a total of more than 300 class averages which were respectively calculated from a total of 6,681 CD147 particles; 5,073 particles of spike(RBD); 12,426 particles of CD147-spike(RBD) complexes. Scale bars: 10 nm. e The co-localization of CD147 and spike protein was observed by immuno-electron microscope. Virions (orange arrows) were observed in virus-infected Vero E6 cells and lung and kidney tissues from COVID-19 patient. The co-localization of CD147 (20 nm-gold colloid, red arrows) and spike protein (10 nm-gold colloid, yellow arrows) in SARS-CoV-2 infected Vero E6 cells and lung and kidney tissues from a patient with COVID-19. Scale bars: 200 nm
    Figure Legend Snippet: Identification of the interaction and co-localization between CD147 and spike protein. a – c The interaction of CD147 and spike was detected by SPR assay ( a ), ELISA ( b ), and Co-IP assay ( c ). The mouse IgG (mIgG) and rabbit IgG (rIgG) were served as negative controls. d OpNS-EM images of CD147, spike(RBD) and CD147-spike(RBD) complexes. Left panels showed the survey view of the micrograph. Right panels showed 8 class averages were selected from a total of more than 300 class averages which were respectively calculated from a total of 6,681 CD147 particles; 5,073 particles of spike(RBD); 12,426 particles of CD147-spike(RBD) complexes. Scale bars: 10 nm. e The co-localization of CD147 and spike protein was observed by immuno-electron microscope. Virions (orange arrows) were observed in virus-infected Vero E6 cells and lung and kidney tissues from COVID-19 patient. The co-localization of CD147 (20 nm-gold colloid, red arrows) and spike protein (10 nm-gold colloid, yellow arrows) in SARS-CoV-2 infected Vero E6 cells and lung and kidney tissues from a patient with COVID-19. Scale bars: 200 nm

    Techniques Used: SPR Assay, Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay, Microscopy, Infection

    SARS-CoV-2 employs the CD147 receptor for host cell entry. a , b Left, Vero E6 and BEAS-2B cells were transfected with shRNA for CD147 gene silence; the gene expression level was detected by real-time PCR. Right, SARS-CoV-2 infection test was performed in Vero E6-shCD147 and BEAS-2B-shCD147 cells. At 48 h after infection, the virus copy number was detected with quantitative PCR (** p
    Figure Legend Snippet: SARS-CoV-2 employs the CD147 receptor for host cell entry. a , b Left, Vero E6 and BEAS-2B cells were transfected with shRNA for CD147 gene silence; the gene expression level was detected by real-time PCR. Right, SARS-CoV-2 infection test was performed in Vero E6-shCD147 and BEAS-2B-shCD147 cells. At 48 h after infection, the virus copy number was detected with quantitative PCR (** p

    Techniques Used: Transfection, shRNA, Expressing, Real-time Polymerase Chain Reaction, Infection

    SARS-CoV-2 invades lung tissues of hCD147 mice and causes pathologic changes. a The WT mice and hCD147 mice were infected with SARS-CoV-2. At 48 h after infection, the viral load of lung tissues was detected with quantitative PCR. b The histopathological changes of lung tissues were detected in WT mice and hCD147 mice by HE staining. Scale bars: 50 μm
    Figure Legend Snippet: SARS-CoV-2 invades lung tissues of hCD147 mice and causes pathologic changes. a The WT mice and hCD147 mice were infected with SARS-CoV-2. At 48 h after infection, the viral load of lung tissues was detected with quantitative PCR. b The histopathological changes of lung tissues were detected in WT mice and hCD147 mice by HE staining. Scale bars: 50 μm

    Techniques Used: Mouse Assay, Infection, Real-time Polymerase Chain Reaction, Staining

    33) Product Images from "Immune response to vaccine candidates based on different types of nanoscaffolded RBD domain of the SARS-CoV-2 spike protein"

    Article Title: Immune response to vaccine candidates based on different types of nanoscaffolded RBD domain of the SARS-CoV-2 spike protein

    Journal: bioRxiv

    doi: 10.1101/2020.08.28.244269

    Neutralization of binding of viral RBD to the ACE2 receptor and inhibition of pseudoviral infection of cells by mouse antisera. A) Sera of mice immunized with DNA vaccines comprising scaffolded RBD were diluted and pre-incubated with Spike protein. Afterwards, Spike that bound to ACE2 was detected using streptactin-HRP. Mean and SEM of 6 (RBD-AaLs) or 5 (all others) biological replicates are shown (A) Sera of mice immunized with DNA vaccines comprising scaffolded RBD were diluted 50-fold and Spike-pseudotyped virus infection of ACE2 and TMPRSS2 –transfected HEK293 cells was followed by luminescence. Mean and SEM of 6 (RBD-AaLs) or 5 (RBD-bann, RBD-foldon-RBD, RBD-ferritin) or 4 (empty pcDNA3 vector, RBD) biological replicates are shown (B). *P
    Figure Legend Snippet: Neutralization of binding of viral RBD to the ACE2 receptor and inhibition of pseudoviral infection of cells by mouse antisera. A) Sera of mice immunized with DNA vaccines comprising scaffolded RBD were diluted and pre-incubated with Spike protein. Afterwards, Spike that bound to ACE2 was detected using streptactin-HRP. Mean and SEM of 6 (RBD-AaLs) or 5 (all others) biological replicates are shown (A) Sera of mice immunized with DNA vaccines comprising scaffolded RBD were diluted 50-fold and Spike-pseudotyped virus infection of ACE2 and TMPRSS2 –transfected HEK293 cells was followed by luminescence. Mean and SEM of 6 (RBD-AaLs) or 5 (RBD-bann, RBD-foldon-RBD, RBD-ferritin) or 4 (empty pcDNA3 vector, RBD) biological replicates are shown (B). *P

    Techniques Used: Neutralization, Binding Assay, Inhibition, Infection, Mouse Assay, Incubation, Transfection, Plasmid Preparation

    Total IgG in mice that underwent switch immunization. Mice were immunized with combinations of differently scaffolded RBD plasmid DNA (β-annulus and foldon) for prime and boost immunization. Titers of antibodies against RBD after prime and boost (A,B) and against Spike protein (C, D) were determined via ELISA. Graphs represent mean of EPT of group of mice (n=6 per group). Each dot represents an individual animal. To determine NS, Mann-Whitney test was performed.
    Figure Legend Snippet: Total IgG in mice that underwent switch immunization. Mice were immunized with combinations of differently scaffolded RBD plasmid DNA (β-annulus and foldon) for prime and boost immunization. Titers of antibodies against RBD after prime and boost (A,B) and against Spike protein (C, D) were determined via ELISA. Graphs represent mean of EPT of group of mice (n=6 per group). Each dot represents an individual animal. To determine NS, Mann-Whitney test was performed.

    Techniques Used: Mouse Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Analysis of different classes of antibodies against RBD for different scaffolded RBDs and immunization by the scaffold. Mice were immunized with different combination of RBD plasmid DNA. End point titers 6 weeks after the first immunization of IgA (A), IgM (B), IgG1 (C), IgG2b (D) and IgG3 (E) against RBD protein were determined by ELISA. Graphs represent mean of EPT of group of mice (n=5 per group). Each dot represents an individual animal.
    Figure Legend Snippet: Analysis of different classes of antibodies against RBD for different scaffolded RBDs and immunization by the scaffold. Mice were immunized with different combination of RBD plasmid DNA. End point titers 6 weeks after the first immunization of IgA (A), IgM (B), IgG1 (C), IgG2b (D) and IgG3 (E) against RBD protein were determined by ELISA. Graphs represent mean of EPT of group of mice (n=5 per group). Each dot represents an individual animal.

    Techniques Used: Mouse Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

    Total IgG against scaffold in mice that underwent switch immunization. Mice were immunized with combinations of differently scaffolded RBD plasmid DNA (β-annulus and foldon) for prime and boost immunization and vice versa. Titers of antibodies against scaffold (depicted in blue) after prime and boost were determined via ELISA. Graphs represent mean of EPT of group of mice (n=6 per group). Each dot represents an individual animal.
    Figure Legend Snippet: Total IgG against scaffold in mice that underwent switch immunization. Mice were immunized with combinations of differently scaffolded RBD plasmid DNA (β-annulus and foldon) for prime and boost immunization and vice versa. Titers of antibodies against scaffold (depicted in blue) after prime and boost were determined via ELISA. Graphs represent mean of EPT of group of mice (n=6 per group). Each dot represents an individual animal.

    Techniques Used: Mouse Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

    Protection of pseudoviral infection by DNA plasmid immunization in a mouse model. Mice were immunized by two injections of plasmids separated by two weeks. After one month hACE2 and TMPRRS was introduced by intranasal plasmid transfection followed by intranasal infection with SARS_CoV-2 S-typed virus (PV). Luminescence based on pseudovirus intranasal infection was measured after 24 hrs (A). Bioluminescence imaging revealing the protective state of immunized animals against pseudovirus infection in animals. Subsequent quantification of bioluminescence average radiance was carried out (B, C). Dashed line represent merging of pictures of mice from the same test group taken separately. Each dot represents an individual animal (pcDNA3 n=4; RBD and RBD-bann n=5). **P
    Figure Legend Snippet: Protection of pseudoviral infection by DNA plasmid immunization in a mouse model. Mice were immunized by two injections of plasmids separated by two weeks. After one month hACE2 and TMPRRS was introduced by intranasal plasmid transfection followed by intranasal infection with SARS_CoV-2 S-typed virus (PV). Luminescence based on pseudovirus intranasal infection was measured after 24 hrs (A). Bioluminescence imaging revealing the protective state of immunized animals against pseudovirus infection in animals. Subsequent quantification of bioluminescence average radiance was carried out (B, C). Dashed line represent merging of pictures of mice from the same test group taken separately. Each dot represents an individual animal (pcDNA3 n=4; RBD and RBD-bann n=5). **P

    Techniques Used: Infection, Plasmid Preparation, Mouse Assay, Transfection, Imaging

    Titer of total IgG antibodies against the RBD and Spike protein for immunization with plasmids for different scaffolded RBDs and scaffold alone. Mice were immunized with different combination of RBD plasmid DNA, complexed with jetPEI- in vivo transfection reagent, according to immunization protocol (A). End point titer (EPT) for total IgG against RBD (B-D) and against Spike protein (E-G). Graphs represent mean of EPT of group of mice (n=5 per group). Each dot represents an individual animal. *P
    Figure Legend Snippet: Titer of total IgG antibodies against the RBD and Spike protein for immunization with plasmids for different scaffolded RBDs and scaffold alone. Mice were immunized with different combination of RBD plasmid DNA, complexed with jetPEI- in vivo transfection reagent, according to immunization protocol (A). End point titer (EPT) for total IgG against RBD (B-D) and against Spike protein (E-G). Graphs represent mean of EPT of group of mice (n=5 per group). Each dot represents an individual animal. *P

    Techniques Used: Mouse Assay, Plasmid Preparation, In Vivo, Transfection

    DNA plasmid immunization with naked DNA. Mice were immunized with 20 μg per animal of naked DNA (empty vector, RBD, RBD-bann), dissolved in 150 mM NaCl. End point titer (EPT) for total IgG against RBD (A) and against Spike protein (B) were determined by ELISA. Graphs represent mean of EPT of group of mice (n=6 per group). Each dot represents an individual animal. *P
    Figure Legend Snippet: DNA plasmid immunization with naked DNA. Mice were immunized with 20 μg per animal of naked DNA (empty vector, RBD, RBD-bann), dissolved in 150 mM NaCl. End point titer (EPT) for total IgG against RBD (A) and against Spike protein (B) were determined by ELISA. Graphs represent mean of EPT of group of mice (n=6 per group). Each dot represents an individual animal. *P

    Techniques Used: Plasmid Preparation, Mouse Assay, Enzyme-linked Immunosorbent Assay

    Secretion of RDB protein domains fused to different scaffolding proteins produced in plasmid-transfected mammalian cells and size analysis of the isolated RBD-bann protein. Supernatant of HEK293 cells transfected with indicated construct was harvested 3 days post transfection and the presence of differently scaffolded RBD domain variants was detected with anti RBD antibodies (A). Size analysis of the purified RBD-bann by DLS confirms the presence of particles around 500 nm (B).
    Figure Legend Snippet: Secretion of RDB protein domains fused to different scaffolding proteins produced in plasmid-transfected mammalian cells and size analysis of the isolated RBD-bann protein. Supernatant of HEK293 cells transfected with indicated construct was harvested 3 days post transfection and the presence of differently scaffolded RBD domain variants was detected with anti RBD antibodies (A). Size analysis of the purified RBD-bann by DLS confirms the presence of particles around 500 nm (B).

    Techniques Used: Scaffolding, Produced, Plasmid Preparation, Transfection, Isolation, Construct, Purification

    34) Product Images from "Human Mesenchymal Stromal Cells Are Resistant to SARS-CoV-2 Infection under Steady-State, Inflammatory Conditions and in the Presence of SARS-CoV-2-Infected Cells"

    Article Title: Human Mesenchymal Stromal Cells Are Resistant to SARS-CoV-2 Infection under Steady-State, Inflammatory Conditions and in the Presence of SARS-CoV-2-Infected Cells

    Journal: Stem Cell Reports

    doi: 10.1016/j.stemcr.2020.09.003

    Evaluation of SARS-CoV-2 Infection of MSCs Evaluation of SARS-CoV-2 infection of MSCs under steady-state and inflammatory conditions and in the presence of SARS-CoV-2-infected Caco-2 cells. SARS-CoV-2 infection is identified by SARS-CoV-2 S protein staining (red). All MSCs and Caco-2 cells experiments were repeated in three independent settings from three BM-MSC donors and three ASC donors, and were performed in three biological replicates each. One representative picture is shown for each condition. (A) Caco-2 cells without SARS-CoV-2; (B) Caco-2 cells with SARS-CoV-2 MOI1; (C) SARS-CoV-2 replication quantified by qPCR detecting high copy numbers in Caco-2 cells infected by SARS-CoV-2; error bars: SD; (D) BM-MSC steady state with SARS-CoV-2 MOI1; (E) ASC steady state with SARS-CoV-2 MOI1; (F) BM-MSC inflammatory conditions with SARS-CoV-2 MOI1; (G) Co-culture BM-MSC:Caco-2 cells (10:1) with SARS-CoV-2 MOI1; BM-MSCs (black star) + Caco-2 cells (white star). Scale bars, 100 μm and 20 μm (inset in G).
    Figure Legend Snippet: Evaluation of SARS-CoV-2 Infection of MSCs Evaluation of SARS-CoV-2 infection of MSCs under steady-state and inflammatory conditions and in the presence of SARS-CoV-2-infected Caco-2 cells. SARS-CoV-2 infection is identified by SARS-CoV-2 S protein staining (red). All MSCs and Caco-2 cells experiments were repeated in three independent settings from three BM-MSC donors and three ASC donors, and were performed in three biological replicates each. One representative picture is shown for each condition. (A) Caco-2 cells without SARS-CoV-2; (B) Caco-2 cells with SARS-CoV-2 MOI1; (C) SARS-CoV-2 replication quantified by qPCR detecting high copy numbers in Caco-2 cells infected by SARS-CoV-2; error bars: SD; (D) BM-MSC steady state with SARS-CoV-2 MOI1; (E) ASC steady state with SARS-CoV-2 MOI1; (F) BM-MSC inflammatory conditions with SARS-CoV-2 MOI1; (G) Co-culture BM-MSC:Caco-2 cells (10:1) with SARS-CoV-2 MOI1; BM-MSCs (black star) + Caco-2 cells (white star). Scale bars, 100 μm and 20 μm (inset in G).

    Techniques Used: Infection, Staining, Real-time Polymerase Chain Reaction, Co-Culture Assay

    35) Product Images from "CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells"

    Article Title: CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells

    Journal: Signal Transduction and Targeted Therapy

    doi: 10.1038/s41392-020-00426-x

    CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p
    Figure Legend Snippet: CD147 is an alternative receptor for SARS-CoV-2 infection in ACE2-deficient cell types. a No interaction of CD147 and ACE2 was detected by Co-IP assay. The mIgG and rIgG were served as negative controls. b No co-localization was found between CD147 and ACE2 by FRET. The color bar denotes FRET ratio. Scale bars: 10 μm. c No co-localization of CD147-ACE2 and the co-localizations of spike-ACE2 and spike-CD147 were observed by immuno-electron microscope (scale bars: 200 nm) and multicolor immunofluorescence (magnification: ×200) in lung tissues from COVID-19 patient. Spike protein, 10 nm-gold colloid, purple arrows; CD147, 20 nm-gold colloid, blue arrows; and ACE2, 40 nm-gold colloid, green arrows. d Virions (red arrows) were observed in lymphocytes of lung tissues from COVID-19 patient. Scale bars: 500 nm. The localization of spike protein and CD3 was analyzed by multicolor immunofluorescence staining. Magnification: ×200. e The gene expressions of CD147 and ACE2 in CD4+ and CD8+ T cells were detected by real-time PCR (*** p

    Techniques Used: Infection, Co-Immunoprecipitation Assay, Microscopy, Immunofluorescence, Multicolor Immunofluorescence Staining, Real-time Polymerase Chain Reaction

    SARS-CoV-2 enters the host cells through CD147-mediated endocytosis. a The sequential endocytosis of SARS-CoV-2 was observed in Vero E6 cells by electron microscope. Scale bars: 200 nm. b The co-localization of spike protein, CD147, and Rab5 were analyzed in BHK-21-CD147 cells and lung tissues from COVID-19 patient by multicolor immunofluorescence staining. Magnification: ×200.
    Figure Legend Snippet: SARS-CoV-2 enters the host cells through CD147-mediated endocytosis. a The sequential endocytosis of SARS-CoV-2 was observed in Vero E6 cells by electron microscope. Scale bars: 200 nm. b The co-localization of spike protein, CD147, and Rab5 were analyzed in BHK-21-CD147 cells and lung tissues from COVID-19 patient by multicolor immunofluorescence staining. Magnification: ×200.

    Techniques Used: Microscopy, Multicolor Immunofluorescence Staining

    Identification of the interaction and co-localization between CD147 and spike protein. a – c The interaction of CD147 and spike was detected by SPR assay ( a ), ELISA ( b ), and Co-IP assay ( c ). The mouse IgG (mIgG) and rabbit IgG (rIgG) were served as negative controls. d OpNS-EM images of CD147, spike(RBD) and CD147-spike(RBD) complexes. Left panels showed the survey view of the micrograph. Right panels showed 8 class averages were selected from a total of more than 300 class averages which were respectively calculated from a total of 6,681 CD147 particles; 5,073 particles of spike(RBD); 12,426 particles of CD147-spike(RBD) complexes. Scale bars: 10 nm. e The co-localization of CD147 and spike protein was observed by immuno-electron microscope. Virions (orange arrows) were observed in virus-infected Vero E6 cells and lung and kidney tissues from COVID-19 patient. The co-localization of CD147 (20 nm-gold colloid, red arrows) and spike protein (10 nm-gold colloid, yellow arrows) in SARS-CoV-2 infected Vero E6 cells and lung and kidney tissues from a patient with COVID-19. Scale bars: 200 nm
    Figure Legend Snippet: Identification of the interaction and co-localization between CD147 and spike protein. a – c The interaction of CD147 and spike was detected by SPR assay ( a ), ELISA ( b ), and Co-IP assay ( c ). The mouse IgG (mIgG) and rabbit IgG (rIgG) were served as negative controls. d OpNS-EM images of CD147, spike(RBD) and CD147-spike(RBD) complexes. Left panels showed the survey view of the micrograph. Right panels showed 8 class averages were selected from a total of more than 300 class averages which were respectively calculated from a total of 6,681 CD147 particles; 5,073 particles of spike(RBD); 12,426 particles of CD147-spike(RBD) complexes. Scale bars: 10 nm. e The co-localization of CD147 and spike protein was observed by immuno-electron microscope. Virions (orange arrows) were observed in virus-infected Vero E6 cells and lung and kidney tissues from COVID-19 patient. The co-localization of CD147 (20 nm-gold colloid, red arrows) and spike protein (10 nm-gold colloid, yellow arrows) in SARS-CoV-2 infected Vero E6 cells and lung and kidney tissues from a patient with COVID-19. Scale bars: 200 nm

    Techniques Used: SPR Assay, Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay, Microscopy, Infection

    SARS-CoV-2 employs the CD147 receptor for host cell entry. a , b Left, Vero E6 and BEAS-2B cells were transfected with shRNA for CD147 gene silence; the gene expression level was detected by real-time PCR. Right, SARS-CoV-2 infection test was performed in Vero E6-shCD147 and BEAS-2B-shCD147 cells. At 48 h after infection, the virus copy number was detected with quantitative PCR (** p
    Figure Legend Snippet: SARS-CoV-2 employs the CD147 receptor for host cell entry. a , b Left, Vero E6 and BEAS-2B cells were transfected with shRNA for CD147 gene silence; the gene expression level was detected by real-time PCR. Right, SARS-CoV-2 infection test was performed in Vero E6-shCD147 and BEAS-2B-shCD147 cells. At 48 h after infection, the virus copy number was detected with quantitative PCR (** p

    Techniques Used: Transfection, shRNA, Expressing, Real-time Polymerase Chain Reaction, Infection

    36) Product Images from "Development of a Synthetic Poxvirus-Based SARS-CoV-2 Vaccine"

    Article Title: Development of a Synthetic Poxvirus-Based SARS-CoV-2 Vaccine

    Journal: bioRxiv

    doi: 10.1101/2020.07.01.183236

    Humoral immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice immunized twice in a three week interval with 5×10 7 PFU of the single and double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific humoral immune responses A-B) Binding antibodies. S, RBD, and N-specific binding antibodies induced by the vaccine vectors were evaluated after the first (A) and second (B) immunization by ELISA. Dashed lines in A and B indicate median binding antibody endpoint titers measured in convalescent human sera (Figure S4). One-way ANOVA with Tukey’s multiple comparison test was used to evaluate differences between binding antibody end-point titers. C) IgG2a/IgG1 isotype ratio. S-, RBD-, and N-specific binding antibodies of the IgG2a and IgG1 isotype were measured after the second immunization using 1:10,000 serum dilution, and absorbance reading was used to calculate IgG2a/IgG1 antibody ratio. One-way ANOVA with Dunnett’s multiple comparison test was used to compare each group mean IgG2a/IgG1 ratio to a ratio of 1 (balanced Th1/Th2 response). D-G) NAb responses. SARS-CoV-2-specific NAb (NT90 titer) induced by the vaccine vectors were measured after the first (D, F) and second (E, G) immunization against SARS-CoV-2 pseudovirus (pv) (D-E) or infectious SARS-CoV-2 virus (F-G) in pooled sera of immunized mice. Shown is the average NT90 measured in duplicate (D-E) or triplicate (F-G) infection. N/A=failed quality control of the samples. Dotted lines indicate lowest antibody dilution included in the analysis. H) SARS-CoV-2/SARS-CoV-2pv correlation analysis. Correlation analysis of NT90 measured in mouse sera after one and two immunizations using infectious SARS-CoV-2 virus and SARS-CoV-2pv. Pearson correlation coefficient (r) was calculated in H. *p
    Figure Legend Snippet: Humoral immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice immunized twice in a three week interval with 5×10 7 PFU of the single and double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific humoral immune responses A-B) Binding antibodies. S, RBD, and N-specific binding antibodies induced by the vaccine vectors were evaluated after the first (A) and second (B) immunization by ELISA. Dashed lines in A and B indicate median binding antibody endpoint titers measured in convalescent human sera (Figure S4). One-way ANOVA with Tukey’s multiple comparison test was used to evaluate differences between binding antibody end-point titers. C) IgG2a/IgG1 isotype ratio. S-, RBD-, and N-specific binding antibodies of the IgG2a and IgG1 isotype were measured after the second immunization using 1:10,000 serum dilution, and absorbance reading was used to calculate IgG2a/IgG1 antibody ratio. One-way ANOVA with Dunnett’s multiple comparison test was used to compare each group mean IgG2a/IgG1 ratio to a ratio of 1 (balanced Th1/Th2 response). D-G) NAb responses. SARS-CoV-2-specific NAb (NT90 titer) induced by the vaccine vectors were measured after the first (D, F) and second (E, G) immunization against SARS-CoV-2 pseudovirus (pv) (D-E) or infectious SARS-CoV-2 virus (F-G) in pooled sera of immunized mice. Shown is the average NT90 measured in duplicate (D-E) or triplicate (F-G) infection. N/A=failed quality control of the samples. Dotted lines indicate lowest antibody dilution included in the analysis. H) SARS-CoV-2/SARS-CoV-2pv correlation analysis. Correlation analysis of NT90 measured in mouse sera after one and two immunizations using infectious SARS-CoV-2 virus and SARS-CoV-2pv. Pearson correlation coefficient (r) was calculated in H. *p

    Techniques Used: Mouse Assay, Recombinant, Derivative Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Infection

    Cellular immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice immunized twice in a three week interval with 5×10 7 PFU of the single or double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific cellular immune responses. Antigen-specific CD8+ ( A and B ) and CD4+ ( C and D ) T cell responses induced by the vaccine vectors after two immunizations were evaluated by flow cytometry for IFN γ , TNFα, IL-4 and IL-10 secretion following ex vivo antigen stimulation using SARS-CoV-2 S and N-specific peptide libraries. Due to technical issues, 1-3 animals/group were not included in the CD4/TNFα analysis in C and D. One-way ANOVA with Tukey’s multiple comparison test was used to compare differences in % of cytokine-specific T-cells between groups. *p
    Figure Legend Snippet: Cellular immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice immunized twice in a three week interval with 5×10 7 PFU of the single or double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific cellular immune responses. Antigen-specific CD8+ ( A and B ) and CD4+ ( C and D ) T cell responses induced by the vaccine vectors after two immunizations were evaluated by flow cytometry for IFN γ , TNFα, IL-4 and IL-10 secretion following ex vivo antigen stimulation using SARS-CoV-2 S and N-specific peptide libraries. Due to technical issues, 1-3 animals/group were not included in the CD4/TNFα analysis in C and D. One-way ANOVA with Tukey’s multiple comparison test was used to compare differences in % of cytokine-specific T-cells between groups. *p

    Techniques Used: Mouse Assay, Recombinant, Derivative Assay, Flow Cytometry, Ex Vivo

    37) Product Images from "Development of a multi-antigenic SARS-CoV-2 vaccine candidate using a synthetic poxvirus platform"

    Article Title: Development of a multi-antigenic SARS-CoV-2 vaccine candidate using a synthetic poxvirus platform

    Journal: Nature Communications

    doi: 10.1038/s41467-020-19819-1

    Humoral immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice ( n = 5) immunized twice in a 3-week interval with 5 × 10 7 PFU of the single and double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific humoral immune responses. a , b Binding antibodies. S-, RBD-, and N-specific binding antibodies induced by the vaccine vectors were evaluated after the first ( a ) and second ( b ) immunization by ELISA. As a comparison, antigen-specific endpoint titers measured in n = 19 plasma samples from SARS-CoV-2 convalescent individuals (Fig. S4 ) were added in a and b . Data are presented as mean values + SD. One-way ANOVA with Tukey’s multiple comparison test was used to evaluate the differences between binding antibody end-point titers. c IgG2a/IgG1 isotype ratio. S-, RBD-, and N-specific binding antibodies of the IgG2a and IgG1 isotype were measured after the second immunization using 1:10,000 serum dilution, and absorbance reading was used to calculate IgG2a/IgG1 antibody ratio. Lines represent median values. One-way ANOVA with Dunnett’s multiple comparison test was used to compare each group mean IgG2a/IgG1 ratio to a ratio of 1 (balanced Th1/Th2 response). d – g NAb responses. SARS-CoV-2-specific NAb (NT90 titer) induced by the vaccine vectors were measured after the first ( d , f ) and second ( e , g ) immunization against SARS-CoV-2 pseudovirus (pv) ( d , e ) or infectious SARS-CoV-2 virus ( f , g ) in pooled sera of immunized mice. Shown is the average NT90 measured in duplicate ( d , e ) or triplicate ( f , g ) infection. N/A = failed quality control of the samples. Dotted lines indicate lowest antibody dilution included in the analysis. h SARS-CoV-2/SARS-CoV-2pv correlation analysis. Correlation analysis of NT90 measured in mouse sera after one and two immunizations using infectious SARS-CoV-2 virus and SARS-CoV-2pv. Pearson correlation coefficient ( r ) was calculated in h ; *0.05
    Figure Legend Snippet: Humoral immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice ( n = 5) immunized twice in a 3-week interval with 5 × 10 7 PFU of the single and double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific humoral immune responses. a , b Binding antibodies. S-, RBD-, and N-specific binding antibodies induced by the vaccine vectors were evaluated after the first ( a ) and second ( b ) immunization by ELISA. As a comparison, antigen-specific endpoint titers measured in n = 19 plasma samples from SARS-CoV-2 convalescent individuals (Fig. S4 ) were added in a and b . Data are presented as mean values + SD. One-way ANOVA with Tukey’s multiple comparison test was used to evaluate the differences between binding antibody end-point titers. c IgG2a/IgG1 isotype ratio. S-, RBD-, and N-specific binding antibodies of the IgG2a and IgG1 isotype were measured after the second immunization using 1:10,000 serum dilution, and absorbance reading was used to calculate IgG2a/IgG1 antibody ratio. Lines represent median values. One-way ANOVA with Dunnett’s multiple comparison test was used to compare each group mean IgG2a/IgG1 ratio to a ratio of 1 (balanced Th1/Th2 response). d – g NAb responses. SARS-CoV-2-specific NAb (NT90 titer) induced by the vaccine vectors were measured after the first ( d , f ) and second ( e , g ) immunization against SARS-CoV-2 pseudovirus (pv) ( d , e ) or infectious SARS-CoV-2 virus ( f , g ) in pooled sera of immunized mice. Shown is the average NT90 measured in duplicate ( d , e ) or triplicate ( f , g ) infection. N/A = failed quality control of the samples. Dotted lines indicate lowest antibody dilution included in the analysis. h SARS-CoV-2/SARS-CoV-2pv correlation analysis. Correlation analysis of NT90 measured in mouse sera after one and two immunizations using infectious SARS-CoV-2 virus and SARS-CoV-2pv. Pearson correlation coefficient ( r ) was calculated in h ; *0.05

    Techniques Used: Mouse Assay, Recombinant, Derivative Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Infection

    Cellular immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice ( n = 5) immunized twice in a 3-week interval with 5 × 10 7 PFU of the single or double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific cellular immune responses. Antigen-specific CD8+ ( a and b ) and CD4+ ( c and d ) T-cell responses induced by the vaccine vectors after two immunizations were evaluated by flow cytometry for IFNγ, TNFα, IL-4, and IL-10 secretion following ex vivo antigen stimulation using SARS-CoV-2 S- and N-specific peptide libraries. Due to technical issues, 1–3 animals/group were not included in the CD4/TNFα analysis in c and d . One-way ANOVA with Tukey’s multiple comparison test was used to compare % of cytokine-specific T cells between immunized mice and mock controls. Lines represent median values; *0.05
    Figure Legend Snippet: Cellular immune responses stimulated by sMVA-CoV2 vectors. Balb/c mice ( n = 5) immunized twice in a 3-week interval with 5 × 10 7 PFU of the single or double recombinant sMVA-CoV2 vectors derived with FPV HP1.441 (sMVA-S/N hp and sMVA-N/S hp) or TROVAC (sMVA-S/N tv, sMVA-N/S tv, sMVA-S tv, sMVA-N tv) were evaluated for SARS-CoV-2-specific cellular immune responses. Antigen-specific CD8+ ( a and b ) and CD4+ ( c and d ) T-cell responses induced by the vaccine vectors after two immunizations were evaluated by flow cytometry for IFNγ, TNFα, IL-4, and IL-10 secretion following ex vivo antigen stimulation using SARS-CoV-2 S- and N-specific peptide libraries. Due to technical issues, 1–3 animals/group were not included in the CD4/TNFα analysis in c and d . One-way ANOVA with Tukey’s multiple comparison test was used to compare % of cytokine-specific T cells between immunized mice and mock controls. Lines represent median values; *0.05

    Techniques Used: Mouse Assay, Recombinant, Derivative Assay, Flow Cytometry, Ex Vivo

    CD8+ T-cell responses induced by sMVA-N/S in HLA transgenic mice. HLA-B*07:02 (B7) transgenic mice ( n = 4) were immunized twice in a 3-week interval with 1 × 10 7 PFU of sMVA-N/S or sMVA control vector (reconstituted with FPV TROVAC). B7 mice ( n = 3) were mock-immunized as additional control. Development of SARS-CoV-2-specific CD8+ T cells was evaluated 1 week post booster immunization. a – d Intracellular cytokine staining. Nucleocapsid- ( a , c ) and Spike-specific ( b , d ) CD8+ T cells were evaluated by intracellular cytokine staining for IFNγ, TNFα, and IL-4 secretion following ex vivo antigen stimulation by N and S peptide libraries, respectively. Panels a and b show the percentage of CD3+/CD8+ T cells secreting IFNγ, TNFα, or IL-4 following peptide stimulation. Panels c and d show relative frequencies of CD8+ T cells secreting one or more cytokines after peptide stimulation. Total percentage of cytokine-secreting cells within CD3+/CD8+ population is indicated under each pie chart. e ELISPOT analysis of IFNγ-secreting cells following stimulation with S and N peptide libraries, S library sub-pools (1S1, 2S1, S2), and N26 peptide containing the HLA-B*07:02-restricted N-specific immunodominant epitope SPRWYFYYL. One-way ANOVA with Dunnett’s multiple comparison test was used in a and b . Two-way ANOVA with Dunnett’s multiple comparison test was used in e . Data in a , b , and e are presented as mean values ± SD; *0.05
    Figure Legend Snippet: CD8+ T-cell responses induced by sMVA-N/S in HLA transgenic mice. HLA-B*07:02 (B7) transgenic mice ( n = 4) were immunized twice in a 3-week interval with 1 × 10 7 PFU of sMVA-N/S or sMVA control vector (reconstituted with FPV TROVAC). B7 mice ( n = 3) were mock-immunized as additional control. Development of SARS-CoV-2-specific CD8+ T cells was evaluated 1 week post booster immunization. a – d Intracellular cytokine staining. Nucleocapsid- ( a , c ) and Spike-specific ( b , d ) CD8+ T cells were evaluated by intracellular cytokine staining for IFNγ, TNFα, and IL-4 secretion following ex vivo antigen stimulation by N and S peptide libraries, respectively. Panels a and b show the percentage of CD3+/CD8+ T cells secreting IFNγ, TNFα, or IL-4 following peptide stimulation. Panels c and d show relative frequencies of CD8+ T cells secreting one or more cytokines after peptide stimulation. Total percentage of cytokine-secreting cells within CD3+/CD8+ population is indicated under each pie chart. e ELISPOT analysis of IFNγ-secreting cells following stimulation with S and N peptide libraries, S library sub-pools (1S1, 2S1, S2), and N26 peptide containing the HLA-B*07:02-restricted N-specific immunodominant epitope SPRWYFYYL. One-way ANOVA with Dunnett’s multiple comparison test was used in a and b . Two-way ANOVA with Dunnett’s multiple comparison test was used in e . Data in a , b , and e are presented as mean values ± SD; *0.05

    Techniques Used: Transgenic Assay, Mouse Assay, Plasmid Preparation, Staining, Ex Vivo, Enzyme-linked Immunospot

    38) Product Images from "Membrane Nanoparticles Derived from ACE2-rich Cells Block SARS-CoV-2 Infection"

    Article Title: Membrane Nanoparticles Derived from ACE2-rich Cells Block SARS-CoV-2 Infection

    Journal: bioRxiv

    doi: 10.1101/2020.08.12.247338

    Inhibition of HEK-293T-hACE2 NPs on SARS-CoV-2 S1. (a) Binding kinetics for NPs and SARS-CoV-2 S1-RBD loaded on SA biosensors. HEK-293T-hACE2 NPs and HEK-293T NPs are 1.1 mg mL −1 in PBS. (b) Western blot determining the content of S1 binding to HK-2 in the absence and presence of NPs. β-actin is the reference. (c) Binding kinetics for increasing concentrations of HEK-293T-hACE2 NPs and SARS-CoV-2 S1-RBD loaded on SA biosensors. (d) Immunofluorescence microscopy observing the adherence of SARS-CoV-2 S1 (green) on HEK-293T-hACE2 NPs. Scale bar indicates 5 μm. (e) Immunofluorescence microscopy revealing the protection of HEK-293T-hACE2 NPs on HK-2 cells exposed to SARS-CoV-2 S1 (green). The region of interest in SARS-CoV-2 S1-treated group is magnified in the embedding graph. Nuclei are stained using DAPI (blue). Scale bar indicates 20 μm.
    Figure Legend Snippet: Inhibition of HEK-293T-hACE2 NPs on SARS-CoV-2 S1. (a) Binding kinetics for NPs and SARS-CoV-2 S1-RBD loaded on SA biosensors. HEK-293T-hACE2 NPs and HEK-293T NPs are 1.1 mg mL −1 in PBS. (b) Western blot determining the content of S1 binding to HK-2 in the absence and presence of NPs. β-actin is the reference. (c) Binding kinetics for increasing concentrations of HEK-293T-hACE2 NPs and SARS-CoV-2 S1-RBD loaded on SA biosensors. (d) Immunofluorescence microscopy observing the adherence of SARS-CoV-2 S1 (green) on HEK-293T-hACE2 NPs. Scale bar indicates 5 μm. (e) Immunofluorescence microscopy revealing the protection of HEK-293T-hACE2 NPs on HK-2 cells exposed to SARS-CoV-2 S1 (green). The region of interest in SARS-CoV-2 S1-treated group is magnified in the embedding graph. Nuclei are stained using DAPI (blue). Scale bar indicates 20 μm.

    Techniques Used: Inhibition, Binding Assay, Western Blot, Immunofluorescence, Microscopy, Staining

    Alleviation of HEK-293T-hACE2 NPs to the metabolic disturbance of HK-2 cells exposed to SARS-CoV-2 S1. (a) Volcano plot showing the changes of cell proteins. The increased proteins VDAC1-3 are labeled. (b) Altered signaling pathways (SPs) in HK-2 exposed to SARS-CoV-2 S1. (c) Heat map displaying the correction of HEK-293T-hACE2 NPs to the imbalanced cell proteins induced by SARS-CoV-2 S1. VDAC1-3 are highlighted in a red frame. (d) Flow cytometry of HK-2 treated with SARS-CoV-2 S1 in the absence and presence of HEK-293T-hACE2 NPs. **, P
    Figure Legend Snippet: Alleviation of HEK-293T-hACE2 NPs to the metabolic disturbance of HK-2 cells exposed to SARS-CoV-2 S1. (a) Volcano plot showing the changes of cell proteins. The increased proteins VDAC1-3 are labeled. (b) Altered signaling pathways (SPs) in HK-2 exposed to SARS-CoV-2 S1. (c) Heat map displaying the correction of HEK-293T-hACE2 NPs to the imbalanced cell proteins induced by SARS-CoV-2 S1. VDAC1-3 are highlighted in a red frame. (d) Flow cytometry of HK-2 treated with SARS-CoV-2 S1 in the absence and presence of HEK-293T-hACE2 NPs. **, P

    Techniques Used: Labeling, Flow Cytometry

    39) Product Images from "Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients"

    Article Title: Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients

    Journal: bioRxiv

    doi: 10.1101/2020.06.15.153064

    Correlation of AIR and ELISA data for SARS-CoV-2 S1+S2 ECD (left) and RBD (right). Exponential trend lines and associated R 2 values are indicated.
    Figure Legend Snippet: Correlation of AIR and ELISA data for SARS-CoV-2 S1+S2 ECD (left) and RBD (right). Exponential trend lines and associated R 2 values are indicated.

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) array exposed to array exposed to 20% FBS + 10% PNHS; (B) array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Angstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.
    Figure Legend Snippet: Response of a commercial anti-SARS-CoV-2 rabbit polyclonal antibody (pAb) on the array. (A) array exposed to array exposed to 20% FBS + 10% PNHS; (B) array exposed to 1 μg/mL anti-SARS-CoV-2 pAb in 20% FBS + 10% PNHS. Strong responses to SARS-CoV-2 S1+S2 ECD, S1, and RBD are observed, as well as smaller cross-reactive responses to HCoV-229E, HCoV-OC43, and MERS spike proteins; (C) quantitative data for the titration. Concentrations of pAb are provided at the top of each column in ng/mL; response values at each concentration for each antigen are provided in Angstroms of build. (D) Titration curves for the four SARS-CoV-2 antigens with standard deviation of replicate probe spots at each concentration.

    Techniques Used: Titration, Concentration Assay, Standard Deviation

    40) Product Images from "SARS-CoV-2 invades host cells via a novel route: CD147-spike protein"

    Article Title: SARS-CoV-2 invades host cells via a novel route: CD147-spike protein

    Journal: bioRxiv

    doi: 10.1101/2020.03.14.988345

    The localization of CD147 and SP observed by immune-electron microscope. (A) The SARS-CoV-2 infected Vero E6 cell with visible SARS-CoV-2 virion (black arrow). ( B and C ) The localization of CD147 (10nm-gold colloid, yellow arrows) and SP (20nm-gold colloid, red arrows) in viral inclusion bodies of SARS-CoV-2 infected Vero E6 cell. Scale bar = 200nm.
    Figure Legend Snippet: The localization of CD147 and SP observed by immune-electron microscope. (A) The SARS-CoV-2 infected Vero E6 cell with visible SARS-CoV-2 virion (black arrow). ( B and C ) The localization of CD147 (10nm-gold colloid, yellow arrows) and SP (20nm-gold colloid, red arrows) in viral inclusion bodies of SARS-CoV-2 infected Vero E6 cell. Scale bar = 200nm.

    Techniques Used: Microscopy, Infection

    Identification of interaction between CD147 and SP. (A) The interaction of CD147 and SP detected by SPR assay, KD = 1.85×10 −7 M. (B) The interaction of CD147 and SP detected by Co-IP assay. Anti-CD147 antibody and anti-SARS-CoV-2 Spike antibody were used for antibody immobilization for Co-IP. The mIgG and rIgG were selected as negative control. (C) The interaction of CD147 and SP detected by ELISA. (D) The ability of Meplazumab to compete with SP for CD147 binding performed by competitive inhibition ELISA.
    Figure Legend Snippet: Identification of interaction between CD147 and SP. (A) The interaction of CD147 and SP detected by SPR assay, KD = 1.85×10 −7 M. (B) The interaction of CD147 and SP detected by Co-IP assay. Anti-CD147 antibody and anti-SARS-CoV-2 Spike antibody were used for antibody immobilization for Co-IP. The mIgG and rIgG were selected as negative control. (C) The interaction of CD147 and SP detected by ELISA. (D) The ability of Meplazumab to compete with SP for CD147 binding performed by competitive inhibition ELISA.

    Techniques Used: SPR Assay, Co-Immunoprecipitation Assay, Negative Control, Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition

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    Western Blot:

    Article Title: Human organs-on-chips as tools for repurposing approved drugs as potential influenza and COVID19 therapeutics in viral pandemics
    Article Snippet: .. Incorporation of the CoV-2 S protein into the CoV2pp was confirmed using Western Blot analysis with anti-CoV2 S1 chimeric monoclonal antibody with combined constant domains of the human IgG1 molecule and mouse variable regions (40150-D001 Sinobiological, 1:500); a recombinant receptor binding domain (RBD) fragment from the S1 region was used as a control (BEI resources, NR-52306). .. Infection assay using pseudotyped viruses in Huh-7 cells Drugs were tested using entry assays for CoV2pp and VSVpp, as previously described .

    Binding Assay:

    Article Title: Human organs-on-chips as tools for repurposing approved drugs as potential influenza and COVID19 therapeutics in viral pandemics
    Article Snippet: .. Incorporation of the CoV-2 S protein into the CoV2pp was confirmed using Western Blot analysis with anti-CoV2 S1 chimeric monoclonal antibody with combined constant domains of the human IgG1 molecule and mouse variable regions (40150-D001 Sinobiological, 1:500); a recombinant receptor binding domain (RBD) fragment from the S1 region was used as a control (BEI resources, NR-52306). .. Infection assay using pseudotyped viruses in Huh-7 cells Drugs were tested using entry assays for CoV2pp and VSVpp, as previously described .

    Immunofluorescence:

    Article Title: Lycorine, a non-nucleoside RNA dependent RNA polymerase inhibitor, as potential treatment for emerging coronavirus infections
    Article Snippet: .. After viral infection, cells were fixed with 4% paraformaldehyde at 24 h post infection (pi) and were analyzed by immunofluorescence staining using the anti-MERS-CoV spike protein, anti-SARS-CoV spike protein, or anti-SARS-CoV-2 nucleocapsid protein primary antibodies (Sino Biological Inc., Beijing, China), Alexa Fluor 488 goat anti-rabbit IgG secondary antibody, and Hoechst 33342 (Molecular Probes/Thermo Fisher Scientific, Waltham, MA, USA). .. Images were acquired by Perkin Elmer Operetta imaging system (20 × ; Waltham, MA, USA).

    Direct ELISA:

    Article Title: Therapeutic antibodies, targeting the SARS-CoV-2 spike N-terminal domain, protect lethally infected K18-hACE2 mice
    Article Snippet: .. ELISA Direct ELISA consisted of coating microtiter plates with 1 μg/ml of recombinant SARS-CoV-2 spike, S1 domain, RBD or NTD subunits. .. For phage ELISA, HRP-conjugated anti-M13 antibody (Sino Biological, USA, Cat# 11973-MM05T-H lot HO13AU601; used at 1:8000 working dilution) was used following detection with TMB substrate (Millipore, USA).

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    Sino Biological anti rbd monoclonal antibody
    Detection of <t>SARS-CoV-2</t> spike (S) protein expression and localization. (A) Schematic illustration of the SARS-CoV-2 full-length spike (S-FL) and mutant S variants. The <t>RBD</t> (receptor binding domain) is in subunit S1; the FP (fusion peptide), HR1 (heptad repeat 1), HR2 (heptad repeat 2), TM (transmembrane domain), and CT (cytoplasmic tail) are in subunit S2. The endoplasmic reticulum retrieval signals (“KxHxx” motif) in the CT domain of S-FL were destroyed in S-Mut protein. The C-terminal 19 amino acids were lacking in S-C19del. (B) Detection of SARS-CoV-2 S expression in HKE293T cells by Western blot using the anti-RBD monoclonal antibody. Cells were transfected with pS-FL, pS-Mut, and pS-C19del plasmids or with an empty vector. (C) Detection of SARS-CoV-2 S subcellular localization in HKE293T cells by confocal microscopy. Cells were grown on glass coverslips for 24 h preceding transfection of plasmids encoding S protein variants. The cells were harvested and labeled with the corresponding antibodies. Calreticulin, ER marker. Nuclei were counterstained with DAPI. Bar = 20 μm.
    Anti Rbd Monoclonal Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 1 article reviews
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    anti rbd monoclonal antibody - by Bioz Stars, 2021-02
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    95
    Sino Biological sars cov 2 s1
    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of <t>SARS-CoV-2</t> S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.
    Sars Cov 2 S1, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 s1/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 s1 - by Bioz Stars, 2021-02
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    Image Search Results


    Detection of SARS-CoV-2 spike (S) protein expression and localization. (A) Schematic illustration of the SARS-CoV-2 full-length spike (S-FL) and mutant S variants. The RBD (receptor binding domain) is in subunit S1; the FP (fusion peptide), HR1 (heptad repeat 1), HR2 (heptad repeat 2), TM (transmembrane domain), and CT (cytoplasmic tail) are in subunit S2. The endoplasmic reticulum retrieval signals (“KxHxx” motif) in the CT domain of S-FL were destroyed in S-Mut protein. The C-terminal 19 amino acids were lacking in S-C19del. (B) Detection of SARS-CoV-2 S expression in HKE293T cells by Western blot using the anti-RBD monoclonal antibody. Cells were transfected with pS-FL, pS-Mut, and pS-C19del plasmids or with an empty vector. (C) Detection of SARS-CoV-2 S subcellular localization in HKE293T cells by confocal microscopy. Cells were grown on glass coverslips for 24 h preceding transfection of plasmids encoding S protein variants. The cells were harvested and labeled with the corresponding antibodies. Calreticulin, ER marker. Nuclei were counterstained with DAPI. Bar = 20 μm.

    Journal: Genes & Diseases

    Article Title: Development of cell-based pseudovirus entry assay to identify potential viral entry inhibitors and neutralizing antibodies against SARS-CoV-2

    doi: 10.1016/j.gendis.2020.07.006

    Figure Lengend Snippet: Detection of SARS-CoV-2 spike (S) protein expression and localization. (A) Schematic illustration of the SARS-CoV-2 full-length spike (S-FL) and mutant S variants. The RBD (receptor binding domain) is in subunit S1; the FP (fusion peptide), HR1 (heptad repeat 1), HR2 (heptad repeat 2), TM (transmembrane domain), and CT (cytoplasmic tail) are in subunit S2. The endoplasmic reticulum retrieval signals (“KxHxx” motif) in the CT domain of S-FL were destroyed in S-Mut protein. The C-terminal 19 amino acids were lacking in S-C19del. (B) Detection of SARS-CoV-2 S expression in HKE293T cells by Western blot using the anti-RBD monoclonal antibody. Cells were transfected with pS-FL, pS-Mut, and pS-C19del plasmids or with an empty vector. (C) Detection of SARS-CoV-2 S subcellular localization in HKE293T cells by confocal microscopy. Cells were grown on glass coverslips for 24 h preceding transfection of plasmids encoding S protein variants. The cells were harvested and labeled with the corresponding antibodies. Calreticulin, ER marker. Nuclei were counterstained with DAPI. Bar = 20 μm.

    Article Snippet: The anti-RBD monoclonal antibody against the SARS-CoV-2 S protein was kindly provided by Prof. Aishun Jin (College of Basic Medial, Chongqing Medical University).

    Techniques: Expressing, Mutagenesis, Binding Assay, Western Blot, Transfection, Plasmid Preparation, Confocal Microscopy, Labeling, Marker

    S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Journal: bioRxiv

    Article Title: Rapid and quantitative detection of COVID-19 markers in micro-liter sized samples

    doi: 10.1101/2020.04.20.052233

    Figure Lengend Snippet: S1 protein detection. (A) Illustration of the assay mechanism. The sample-to-answer time of this assay is 20 minutes. (B) Entire dynamic ranges of SARS-CoV-2 S1 protein (red squares) and SARS-CoV S1 protein (black circles) in 10 times diluted human serum. The averaged background is subtracted from all data points. The solid lines are the linear fit of the data in the log-log scale. The grey shaded area marks 3×standard deviation of the background. The lower limit of detection (LLOD) for SARS-CoV-2 S1 protein and SARS-CoV S1 is 0.4 ng/mL and 0.2 ng/mL, respectively. (C) Calibration curves for S1 proteins between 0.78 and 200 ng/mL. The error bars are generated from duplicate measurements.

    Article Snippet: Conversely, D006’s binding affinity towards SARS-CoV S1 is weaker than SARS-CoV-2 S1.

    Techniques: Generated