mouse igg1 fc domain  (Sino Biological)


Bioz Verified Symbol Sino Biological is a verified supplier
Bioz Manufacturer Symbol Sino Biological manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Name:
    SARS CoV 2 2019 nCoV Spike RBD mFc Recombinant Protein HPLC verified COVID 19 Spike RBD Research
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with the Fc region of mouse IgG1 at the C terminus
    Catalog Number:
    40592-v05h
    Price:
    None
    Category:
    recombinant protein
    Product Aliases:
    coronavirus spike Protein 2019-nCoV, cov spike Protein 2019-nCoV, ncov RBD Protein 2019-nCoV, ncov s1 Protein 2019-nCoV, ncov s2 Protein 2019-nCoV, ncov spike Protein 2019-nCoV, NCP-CoV RBD Protein 2019-nCoV, NCP-CoV s1 Protein 2019-nCoV, NCP-CoV s2 Protein 2019-nCoV, NCP-CoV Spike Protein 2019-nCoV, novel coronavirus RBD Protein 2019-nCoV, novel coronavirus s1 Protein 2019-nCoV, novel coronavirus s2 Protein 2019-nCoV, novel coronavirus spike Protein 2019-nCoV, RBD Protein 2019-nCoV, S1 Protein 2019-nCoV, S2 Protein 2019-nCoV, Spike RBD Protein 2019-nCoV
    Host:
    HEK293 Cells
    Buy from Supplier


    Structured Review

    Sino Biological mouse igg1 fc domain
    Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human <t>IgG</t> secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with the Fc region of mouse IgG1 at the C terminus
    https://www.bioz.com/result/mouse igg1 fc domain/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse igg1 fc domain - by Bioz Stars, 2021-03
    94/100 stars

    Images

    1) Product Images from "Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein"

    Article Title: Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein

    Journal: bioRxiv

    doi: 10.1101/2020.05.12.091462

    Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human IgG secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.
    Figure Legend Snippet: Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human IgG secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.

    Techniques Used: Functional Assay, Binding Assay, Fluorescence, Clone Assay

    2) Product Images from "The efficacy assessment of convalescent plasma therapy for COVID-19 patients: a multi-center case series"

    Article Title: The efficacy assessment of convalescent plasma therapy for COVID-19 patients: a multi-center case series

    Journal: Signal Transduction and Targeted Therapy

    doi: 10.1038/s41392-020-00329-x

    Changes of laboratory results before and at day 1–5 after convalescent plasma transfusion. a , b SARS-CoV-2 specific IgG and IgM levels, respectively, determined by MCLIA. c , d Cycle threshold (Ct) values of ORF1ab-gene and N-gene, respectively. A Ct value of 40 was defined as undetectable. e PaO 2 /FiO 2 (normal range: 400–500 mmHg). f White blood cell count (normal range: 3.5–9.5). g Lymphocyte count (normal range: 1.1–3.2). h C-reactive protein (normal range:
    Figure Legend Snippet: Changes of laboratory results before and at day 1–5 after convalescent plasma transfusion. a , b SARS-CoV-2 specific IgG and IgM levels, respectively, determined by MCLIA. c , d Cycle threshold (Ct) values of ORF1ab-gene and N-gene, respectively. A Ct value of 40 was defined as undetectable. e PaO 2 /FiO 2 (normal range: 400–500 mmHg). f White blood cell count (normal range: 3.5–9.5). g Lymphocyte count (normal range: 1.1–3.2). h C-reactive protein (normal range:

    Techniques Used: Cell Counting

    SARS-CoV-2 specific antibody levels of CP samples measured by serology tests, receptor-binding assay, and pseudovirus based neutralization assay. a The correlations among anti-SARS-CoV-2 specific IgG and IgM titers detected by commercial MCLIA kits, anti-S-RBD and anti-NP specific IgG titers determined by in-house ELISA assays, inhibition activity measured by a receptor-binding assay, and neutralizing antibody titer measured by a pseudovirus based neutralization assay. b Comparisons of antibody levels between CP samples collected before and after 21 days from symptom onset. MCLIA magnetic chemiluminescence enzyme immunoassay, ELISA enzyme-linked immunosorbent assay, RBD receptor binding domains, NP nucleoprotein, IT50 inhibitory titer which was calculated with the dilution of plasma that inhibits 50% RBD-Fc binding to receptor ACE2, NAT50 neutralizing antibody titer which was calculated with the highest dilution of plasma that resulted in a 50% reduction of virus infection, GMT geometric mean titer, CI confidence interval
    Figure Legend Snippet: SARS-CoV-2 specific antibody levels of CP samples measured by serology tests, receptor-binding assay, and pseudovirus based neutralization assay. a The correlations among anti-SARS-CoV-2 specific IgG and IgM titers detected by commercial MCLIA kits, anti-S-RBD and anti-NP specific IgG titers determined by in-house ELISA assays, inhibition activity measured by a receptor-binding assay, and neutralizing antibody titer measured by a pseudovirus based neutralization assay. b Comparisons of antibody levels between CP samples collected before and after 21 days from symptom onset. MCLIA magnetic chemiluminescence enzyme immunoassay, ELISA enzyme-linked immunosorbent assay, RBD receptor binding domains, NP nucleoprotein, IT50 inhibitory titer which was calculated with the dilution of plasma that inhibits 50% RBD-Fc binding to receptor ACE2, NAT50 neutralizing antibody titer which was calculated with the highest dilution of plasma that resulted in a 50% reduction of virus infection, GMT geometric mean titer, CI confidence interval

    Techniques Used: Reporter Assay, Neutralization, Enzyme-linked Immunosorbent Assay, Inhibition, Activity Assay, Binding Assay, Infection

    3) Product Images from "Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells"

    Article Title: Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells

    Journal: Emerging Microbes & Infections

    doi: 10.1080/22221751.2020.1815589

    Validation of the VSVdG-SARS-CoV-2-Sdel18 pseudovirus assay. (A) Specificity of the pseudovirus assay. A negative sample panel including 59 human sera and 58 mouse sera were used to determine the specificity of this assay. (B) Reproducibility of the pseudovirus assay. One COVID-19 convalescent patient serum sample was tested 14 times on individual plates in three independent experiments. The virus titer of VSVdG-SARS-CoV-2-Sdel18 pseudovirus was consistent in these assays (MOI=0.05). (C) The correlation of neutralizing titer measured by the VSVdG-SARS-CoV-2-Sdel18 pseudovirus assay (ID50, log10) and the wild type SARS-CoV-2 neutralization assay (ID100, log10).
    Figure Legend Snippet: Validation of the VSVdG-SARS-CoV-2-Sdel18 pseudovirus assay. (A) Specificity of the pseudovirus assay. A negative sample panel including 59 human sera and 58 mouse sera were used to determine the specificity of this assay. (B) Reproducibility of the pseudovirus assay. One COVID-19 convalescent patient serum sample was tested 14 times on individual plates in three independent experiments. The virus titer of VSVdG-SARS-CoV-2-Sdel18 pseudovirus was consistent in these assays (MOI=0.05). (C) The correlation of neutralizing titer measured by the VSVdG-SARS-CoV-2-Sdel18 pseudovirus assay (ID50, log10) and the wild type SARS-CoV-2 neutralization assay (ID100, log10).

    Techniques Used: Neutralization

    4) Product Images from "Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19"

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19

    Journal: Signal Transduction and Targeted Therapy

    doi: 10.1038/s41392-020-00301-9

    Neutralizing antibody responses to SARS-CoV-2 in COVID-19 recovered patients. a Scores showing the COVID-19 patient serum-mediated inhibition of the SARS-CoV-2 RBD protein binding to ACE2 protein by ELISA. b Pie charts showing the proportions of patients with high ( > 50, green) or low (
    Figure Legend Snippet: Neutralizing antibody responses to SARS-CoV-2 in COVID-19 recovered patients. a Scores showing the COVID-19 patient serum-mediated inhibition of the SARS-CoV-2 RBD protein binding to ACE2 protein by ELISA. b Pie charts showing the proportions of patients with high ( > 50, green) or low (

    Techniques Used: Inhibition, Protein Binding, Enzyme-linked Immunosorbent Assay

    Subtypes of neutralizing antibodies to SARS-CoV-2 S proteins in COVID-19 recovered patients. a Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by patient sera (no depletion) or S1 antibody-depleted sera (S1-Abs depletion) or S2 antibody-depleted sera (S2-Abs depletion). The dashed line indicates the cutoff value (6.749) determined by the ROC curve analysis. HC healthy control, NC negative control. b , c Pie charts showing the proportions of patients with different neutralizing antibody (NAb) subtype responses in the total 25 patients ( b ), 8 severe patients ( c , left panel), and 17 moderate and mild patients ( c , right panel) of pseudovirus neutralization positive. d Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by “S1-NAbs only” patient sera with RBD antibody depletion (RBD-Abs depletion) or without RBD antibody depletion (no depletion). The dashed line indicates the cutoff value (6.034) determined by the ROC curve analysis. HC healthy control, NC negative control. e Pie chart showing the proportions of “S1-NAbs only” patients with RBD-Nab-dependent or -independent antibody response. Error bars in a , d indicate SEM
    Figure Legend Snippet: Subtypes of neutralizing antibodies to SARS-CoV-2 S proteins in COVID-19 recovered patients. a Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by patient sera (no depletion) or S1 antibody-depleted sera (S1-Abs depletion) or S2 antibody-depleted sera (S2-Abs depletion). The dashed line indicates the cutoff value (6.749) determined by the ROC curve analysis. HC healthy control, NC negative control. b , c Pie charts showing the proportions of patients with different neutralizing antibody (NAb) subtype responses in the total 25 patients ( b ), 8 severe patients ( c , left panel), and 17 moderate and mild patients ( c , right panel) of pseudovirus neutralization positive. d Blocking of luciferase-encoding SARS-CoV-2 typed pseudovirus into ACE2/293T cells by “S1-NAbs only” patient sera with RBD antibody depletion (RBD-Abs depletion) or without RBD antibody depletion (no depletion). The dashed line indicates the cutoff value (6.034) determined by the ROC curve analysis. HC healthy control, NC negative control. e Pie chart showing the proportions of “S1-NAbs only” patients with RBD-Nab-dependent or -independent antibody response. Error bars in a , d indicate SEM

    Techniques Used: Blocking Assay, Luciferase, Negative Control, Neutralization

    Antibody responses to SARS-CoV-2 in COVID-19 recovered patients with different symptom severity. a – c ELISA binding assays of 100-fold diluted COVID-19 patient sera to ELISA plates after coating with SARS-CoV-2 S1 ( a ), RBD ( b ), and S2 ( c ) proteins. The dashed lines in a – c represent the average values of the healthy control groups. * P
    Figure Legend Snippet: Antibody responses to SARS-CoV-2 in COVID-19 recovered patients with different symptom severity. a – c ELISA binding assays of 100-fold diluted COVID-19 patient sera to ELISA plates after coating with SARS-CoV-2 S1 ( a ), RBD ( b ), and S2 ( c ) proteins. The dashed lines in a – c represent the average values of the healthy control groups. * P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Binding Assay

    5) Product Images from "Isolation of and Characterization of Neutralizing Antibodies to Covid-19 from a Large Human Naïve scFv Phage Display Library"

    Article Title: Isolation of and Characterization of Neutralizing Antibodies to Covid-19 from a Large Human Naïve scFv Phage Display Library

    Journal: bioRxiv

    doi: 10.1101/2020.05.19.104281

    FACS examination of RBD-binding antibodies to Covid-19 membrane spike on ID8 cells. Soluble scFv-huFc of 22 hits that were RBD ELISA-positive were incubated with equal amount of ID8 cells, followed by goat anti-human Fc (PE conjugated). Stained cells were analyzed to draw dot-plot by FACS. P2 Gating was set based on the background of secondary antibody staining and considered positive. Only clones having positive percentages were shown. Individual clone was labeled in each plot.
    Figure Legend Snippet: FACS examination of RBD-binding antibodies to Covid-19 membrane spike on ID8 cells. Soluble scFv-huFc of 22 hits that were RBD ELISA-positive were incubated with equal amount of ID8 cells, followed by goat anti-human Fc (PE conjugated). Stained cells were analyzed to draw dot-plot by FACS. P2 Gating was set based on the background of secondary antibody staining and considered positive. Only clones having positive percentages were shown. Individual clone was labeled in each plot.

    Techniques Used: FACS, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation, Staining, Clone Assay, Labeling

    MFI change of ACE2 binding to ID8 cells in the presence of serial diluted RBD-antibodies. 8 Covid-19 spike positive antibody binders (1B1, 1B11, 5C2, 1A12, 1A5, 1G6, 1C10 and 1H2) and one isotype negative control antibody (iso-ctrl in the figure) were mixed with ID8 cells at serial diluted concentrations (10 μg/ml down to 0.31 μg/ml, 2 times dilution) before adding of human ACE2-mFc, which has strong interaction with ID8. MFIs (shown in vertical axis) of individual antibody/concentrations cell group (horizonal axis) were complied.
    Figure Legend Snippet: MFI change of ACE2 binding to ID8 cells in the presence of serial diluted RBD-antibodies. 8 Covid-19 spike positive antibody binders (1B1, 1B11, 5C2, 1A12, 1A5, 1G6, 1C10 and 1H2) and one isotype negative control antibody (iso-ctrl in the figure) were mixed with ID8 cells at serial diluted concentrations (10 μg/ml down to 0.31 μg/ml, 2 times dilution) before adding of human ACE2-mFc, which has strong interaction with ID8. MFIs (shown in vertical axis) of individual antibody/concentrations cell group (horizonal axis) were complied.

    Techniques Used: Binding Assay, Negative Control

    BLITZ epitope mapping of Covid-19 RBD-hits antibodies (A). Rotational mutual interaction examination of the six hits (1B11, 1A5, 1G6, 1C10, 1A12 and 1H2) was captured by protein A sensors, followed by RBD binding and one of the six antibodies plus ACE2. RBD was bound well by individual antibodies but not concurrent binding. (B). 1B1 concurrently bound to RBD with any one of the six other antibodies (1B11, 1A5, 1G6, 1C10, 1A12 and 1H2). (C). 5C2 concurrently bound to RBD with any one of the six other antibodies (1B11, 1A5, 1G6, 1C10, A12 and 1H2). (D). 1B1 and 5C2 mutually competed in binding to RBD
    Figure Legend Snippet: BLITZ epitope mapping of Covid-19 RBD-hits antibodies (A). Rotational mutual interaction examination of the six hits (1B11, 1A5, 1G6, 1C10, 1A12 and 1H2) was captured by protein A sensors, followed by RBD binding and one of the six antibodies plus ACE2. RBD was bound well by individual antibodies but not concurrent binding. (B). 1B1 concurrently bound to RBD with any one of the six other antibodies (1B11, 1A5, 1G6, 1C10, 1A12 and 1H2). (C). 5C2 concurrently bound to RBD with any one of the six other antibodies (1B11, 1A5, 1G6, 1C10, A12 and 1H2). (D). 1B1 and 5C2 mutually competed in binding to RBD

    Techniques Used: Binding Assay

    Venn map of Covid-19 hits and ACE2 on RBD Epitope locations on RBD by the isolated 8 hit antibodies were deduced from the above Blitz results and largely categorized into 3 groups (I, II and III). 1B1 and 5C2 have large overlapped sites in RBD and may block the most part of RBD-ACE2 interaction interface. The group III hits (1B11, 1A5, 1G6, 1C10, 1A12 and 1H2) binds to an independent site from 1B1 or 5C2, however may block a minor side of ACE2-RBD interaction interface.
    Figure Legend Snippet: Venn map of Covid-19 hits and ACE2 on RBD Epitope locations on RBD by the isolated 8 hit antibodies were deduced from the above Blitz results and largely categorized into 3 groups (I, II and III). 1B1 and 5C2 have large overlapped sites in RBD and may block the most part of RBD-ACE2 interaction interface. The group III hits (1B11, 1A5, 1G6, 1C10, 1A12 and 1H2) binds to an independent site from 1B1 or 5C2, however may block a minor side of ACE2-RBD interaction interface.

    Techniques Used: Isolation, Blocking Assay

    Validation of Covid-19 spike expression cell line A mammalian cell line ID8 was generated by transfecting 293FT with a pseudo virus that contains full-length Covid-19 S protein, a transmem-brane motif (TM) and a C-terminal 3xFLAG tag. A . The cartoon illustration of the ID8 cells and membrane anchoring of Covid-19 spike (green), C-terminal FLAG (yellow) and binding of ACE2-mFc (blue) to S trimer. B . FACS plot of intracellular staining of FLAG with anti-FLAG-FITC. C . FACS plot of ID8 cell membrane sequential staining of ACE2-mFc, anti-mouse FITC.
    Figure Legend Snippet: Validation of Covid-19 spike expression cell line A mammalian cell line ID8 was generated by transfecting 293FT with a pseudo virus that contains full-length Covid-19 S protein, a transmem-brane motif (TM) and a C-terminal 3xFLAG tag. A . The cartoon illustration of the ID8 cells and membrane anchoring of Covid-19 spike (green), C-terminal FLAG (yellow) and binding of ACE2-mFc (blue) to S trimer. B . FACS plot of intracellular staining of FLAG with anti-FLAG-FITC. C . FACS plot of ID8 cell membrane sequential staining of ACE2-mFc, anti-mouse FITC.

    Techniques Used: Expressing, Generated, Binding Assay, FACS, Staining

    Monophage ELISA screening Covid-19 RBD phage antibodies EHL library was panned with Covid-19 for 2 rounds. Single colonies form 2 nd -round output was picked to prepare monoclonal phage antibody solutions, examined by ELISA with RBD, mouse Fc and 293F cells. Only RBD positive hits were resorted to DNA sequencing of scFv genes. A . Chart of absorbance (A450) on RBD plate ELISA. B . Amino acid sequence multi-alignment of unique scFv clones positive for RBD binding.
    Figure Legend Snippet: Monophage ELISA screening Covid-19 RBD phage antibodies EHL library was panned with Covid-19 for 2 rounds. Single colonies form 2 nd -round output was picked to prepare monoclonal phage antibody solutions, examined by ELISA with RBD, mouse Fc and 293F cells. Only RBD positive hits were resorted to DNA sequencing of scFv genes. A . Chart of absorbance (A450) on RBD plate ELISA. B . Amino acid sequence multi-alignment of unique scFv clones positive for RBD binding.

    Techniques Used: Enzyme-linked Immunosorbent Assay, DNA Sequencing, Sequencing, Clone Assay, Binding Assay

    Cross-reaction examination of Covid-19 RBD hits by ELISA Binding of 8 COVid-19 RBD-hits was examined by ELISA to the spike proteins (named as SARS-S, MERS-S and Covid-19-S) for SARS-COV-1, MERS-COV and SARS-COV-2 (Covid-19) and measured by absorbance (A450). All hits demonstrated strong positive to Covid-19-S (yellow bars) and none cross-reacted to the other two spikes (green and blue bars).
    Figure Legend Snippet: Cross-reaction examination of Covid-19 RBD hits by ELISA Binding of 8 COVid-19 RBD-hits was examined by ELISA to the spike proteins (named as SARS-S, MERS-S and Covid-19-S) for SARS-COV-1, MERS-COV and SARS-COV-2 (Covid-19) and measured by absorbance (A450). All hits demonstrated strong positive to Covid-19-S (yellow bars) and none cross-reacted to the other two spikes (green and blue bars).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Binding Assay

    Dual binding examination of Covid-19 RBD-hit antibodies All 8 RBD hits separately were incubated together and ACE2-mFc at around equal concentrations before adding to ID8 cells. Both antibody binding and ACE2 binding were detected by corresponding specific secondary antibodies (different fluorescent conjugation). Upper row from left: FITC-conjugated secondary control, 1A5, 1H2, 1C10, 5C2. Lower row from left: ACE2-mFc (alone, positive control), 1A12, 1G6, 1B11 and 1B1
    Figure Legend Snippet: Dual binding examination of Covid-19 RBD-hit antibodies All 8 RBD hits separately were incubated together and ACE2-mFc at around equal concentrations before adding to ID8 cells. Both antibody binding and ACE2 binding were detected by corresponding specific secondary antibodies (different fluorescent conjugation). Upper row from left: FITC-conjugated secondary control, 1A5, 1H2, 1C10, 5C2. Lower row from left: ACE2-mFc (alone, positive control), 1A12, 1G6, 1B11 and 1B1

    Techniques Used: Binding Assay, Incubation, Conjugation Assay, Positive Control

    Related Articles

    Recombinant:

    Article Title: The efficacy assessment of convalescent plasma therapy for COVID-19 patients: a multi-center case series
    Article Snippet: .. 80 ng/ml recombinant SARS-CoV-2 Spike RBD-mFc (Sino Biological) was mixed with the presence or absence of serially diluted CP or serum samples 1:1 and incubated at 37 °C for 1 h, then add the 100 μl mixed solution to the wells. .. Incubated at 37 °C for 10 min, 100 μl of the HRP conjugated goat anti-mouse IgG (ZSGB-BIO) was added to the wells.

    Incubation:

    Article Title: The efficacy assessment of convalescent plasma therapy for COVID-19 patients: a multi-center case series
    Article Snippet: .. 80 ng/ml recombinant SARS-CoV-2 Spike RBD-mFc (Sino Biological) was mixed with the presence or absence of serially diluted CP or serum samples 1:1 and incubated at 37 °C for 1 h, then add the 100 μl mixed solution to the wells. .. Incubated at 37 °C for 10 min, 100 μl of the HRP conjugated goat anti-mouse IgG (ZSGB-BIO) was added to the wells.

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: ELISA-based RBD–ACE2-binding inhibition assay As previously described, 200 ng of ACE2 protein (Sino Biological, 10108-H08H) in 100 μl PBS per well was coated on ELISA plates overnight at 4 °C. .. The ELISA plates were blocked for 1 h with 100 μl blocking buffer (5% FBS and 0.1% Tween 20 in PBS); meanwhile, 50 μl 10-fold diluted patient or healthy control sera were incubated with 7.5 ng SARS-CoV-2 RBD-mouse FC protein (Sino Biological, 40592-V05H) in 50 μl blocking buffer for 1 h. Then the incubated sera/SARS-CoV-2 RBD-mouse FC protein mixture was added into the ELISA plates and allowed to develop for 30 min, followed by PBST washing and incubation with anti-mouse FC HRP antibody (Thermo Fisher Scientific, A16084) for 30 min. Next, the ELISA plates were washed with PBST and treated with TMB buffer (Beyotime, P0209). .. After 5 min, the ELISA reaction was stopped by 1 M HCl stop buffer and determined at 450 nm.

    other:

    Article Title: Human H-ferritin presenting RBM of spike glycoprotein as potential vaccine of SARS-CoV-2
    Article Snippet: MaterialsHuman ACE2 (his tag) (10108-H08H), ACE2 rabbit Antibody (10108-RP01), goat anti-Rabbit/HRP secondary antibody (SSA004, and SARS-CoV-2 (2019-nCoV) and RBD of spike glycoprotein (mFC tag)(40592-V05H) were purchased from Sino Biological (Beijing, China).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: ELISA-based RBD–ACE2-binding inhibition assay As previously described, 200 ng of ACE2 protein (Sino Biological, 10108-H08H) in 100 μl PBS per well was coated on ELISA plates overnight at 4 °C. .. The ELISA plates were blocked for 1 h with 100 μl blocking buffer (5% FBS and 0.1% Tween 20 in PBS); meanwhile, 50 μl 10-fold diluted patient or healthy control sera were incubated with 7.5 ng SARS-CoV-2 RBD-mouse FC protein (Sino Biological, 40592-V05H) in 50 μl blocking buffer for 1 h. Then the incubated sera/SARS-CoV-2 RBD-mouse FC protein mixture was added into the ELISA plates and allowed to develop for 30 min, followed by PBST washing and incubation with anti-mouse FC HRP antibody (Thermo Fisher Scientific, A16084) for 30 min. Next, the ELISA plates were washed with PBST and treated with TMB buffer (Beyotime, P0209). .. After 5 min, the ELISA reaction was stopped by 1 M HCl stop buffer and determined at 450 nm.

    Article Title: Clinical Utility of a Highly Sensitive Lateral Flow Immunoassay as determined by Titer Analysis for the Detection of anti-SARS-CoV-2 Antibodies at the Point-of-Care
    Article Snippet: .. ELISA plates (Immulon 1B, ThermoFisher cat# 3355) 117 were coated overnight at 4°C with 2 μg/mL SARS-CoV-2 spike RBD-mFc tag (Sino Biological 118 cat# 40592-V05H) produced in HEK293 cells. .. Serum samples were incubated on antigen 119 coated plates at 10-fold dilutions ranging from 1:2 to 1:20,000 for 2 hr at room temp.

    Blocking Assay:

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: ELISA-based RBD–ACE2-binding inhibition assay As previously described, 200 ng of ACE2 protein (Sino Biological, 10108-H08H) in 100 μl PBS per well was coated on ELISA plates overnight at 4 °C. .. The ELISA plates were blocked for 1 h with 100 μl blocking buffer (5% FBS and 0.1% Tween 20 in PBS); meanwhile, 50 μl 10-fold diluted patient or healthy control sera were incubated with 7.5 ng SARS-CoV-2 RBD-mouse FC protein (Sino Biological, 40592-V05H) in 50 μl blocking buffer for 1 h. Then the incubated sera/SARS-CoV-2 RBD-mouse FC protein mixture was added into the ELISA plates and allowed to develop for 30 min, followed by PBST washing and incubation with anti-mouse FC HRP antibody (Thermo Fisher Scientific, A16084) for 30 min. Next, the ELISA plates were washed with PBST and treated with TMB buffer (Beyotime, P0209). .. After 5 min, the ELISA reaction was stopped by 1 M HCl stop buffer and determined at 450 nm.

    Produced:

    Article Title: Clinical Utility of a Highly Sensitive Lateral Flow Immunoassay as determined by Titer Analysis for the Detection of anti-SARS-CoV-2 Antibodies at the Point-of-Care
    Article Snippet: .. ELISA plates (Immulon 1B, ThermoFisher cat# 3355) 117 were coated overnight at 4°C with 2 μg/mL SARS-CoV-2 spike RBD-mFc tag (Sino Biological 118 cat# 40592-V05H) produced in HEK293 cells. .. Serum samples were incubated on antigen 119 coated plates at 10-fold dilutions ranging from 1:2 to 1:20,000 for 2 hr at room temp.

    Generated:

    Article Title: Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells
    Article Snippet: Transposon vectors (SBI System Biosciences, PB514B-2) containing the hACE2 gene and transposase plasmids were co-transfected into BHK21 cells and then selected through puromycin resistance and red fluorescence. .. SARS-CoV-2 spike-specific antibody was generated through the immunization of Balb/c mice with SARS-CoV-2 spike protein (RBD, Fc Tag, 40592-V05H, Sino Biological Inc.). .. MAbs against the SARS-CoV-2 RBD were produced with hybridoma technology and were characterized by pseudotyped virus assay.

    Mouse Assay:

    Article Title: Robust neutralization assay based on SARS-CoV-2 S-protein-bearing vesicular stomatitis virus (VSV) pseudovirus and ACE2-overexpressing BHK21 cells
    Article Snippet: Transposon vectors (SBI System Biosciences, PB514B-2) containing the hACE2 gene and transposase plasmids were co-transfected into BHK21 cells and then selected through puromycin resistance and red fluorescence. .. SARS-CoV-2 spike-specific antibody was generated through the immunization of Balb/c mice with SARS-CoV-2 spike protein (RBD, Fc Tag, 40592-V05H, Sino Biological Inc.). .. MAbs against the SARS-CoV-2 RBD were produced with hybridoma technology and were characterized by pseudotyped virus assay.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Sino Biological sars cov 2 antibody
    Viral load of the <t>SARS‐CoV‐2‐infected</t> rhesus macaque model. A. Average viral loads of swabs from the younger group (YG, n = 3, red line) monkeys. B. Average viral load of swabs from the elder group (EG, n = 2, blue line) monkeys. Viral loads of nasal, throat, and anal swab specimens collected from the inoculated macaques on 0, 3, 5, 7, 9, 11, and 14 dpi. C. Viral loads in varies lobe of lung tissue from YG and EG monkeys at day 7 post‐inoculation. RNA was extracted and viral load was determined by qRT‐PCR. All data are presented as mean ± SEM
    Sars Cov 2 Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 antibody/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 antibody - by Bioz Stars, 2021-03
    93/100 stars
      Buy from Supplier

    94
    Sino Biological sars cov 2 2019 ncov spike rbd antibody rabbit pab
    JIB-04 exhibits distinct post-entry antiviral mechanisms (A) Drug combination dose-response matrix and <t>VSV-SARS-CoV-2</t> replication. MA104 cells were treated with JIB-04 and chloroquine or JIB-04 and NTZ for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. (B) Time of compound addition and VSV-SARS-CoV-2 replication. MA104 cells were treated with NTZ or JIB-04 (10 μM) at indicated time points relative to VSV-SARS-CoV-2 infection (MOI=3, 0 hpi). GFP signals at 8 hpi were quantified to calculate the percentage of inhibition. (C) Intracellular SARS-CoV-2 S RNA levels with JIB-04 treatment. MA104 cells were treated with JIB-04 (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1) for 1, 3, 5, and 7 hr. S RNA levels were measured by RT-qPCR. (D) Western blot analysis of SARS-CoV-2 S protein levels with JIB-04 treatment. MA104 cells were treated with JIB-04 (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1) for 1, 3, 5, and 7 hr. FL: full-length. S2: cleaved S2 fragment. (* non-specific band) (E) Histone demethylase siRNA knockdown and RV replication. HEK293 cells were transfected with scrambled siRNA or siRNA targeting indicated histone demethylases for 48 hr and infected with porcine RV (MOI=0.01). Viral RNA copy numbers at 12 hpi were quantified by RT-qPCR. (F) Volcano plot of differentially expressed transcripts with JIB-04 treatment and RV infection. HEK293 cells were treated with DMSO or JIB-04 (10 μM) for 12 hr, and mock-infected (left panel) or infected with porcine RV (MOI=0.01, right panel) for another 12 hr. Red dots represent upregulated genes and green dots represent downregulated genes in JIB-04 treated cells. (G) Expression of three top genes in (F) with JIB-04 treatment. HEK293 cells were treated with JIB-04 (10 μM) for 12 hr and mock-infected or infected porcine RV (MOI=0.01) for 12 hr. mRNA levels of CYP1A1, CYP1B1, and AHRR at 12 hpi were measured by RT-qPCR. (H) Dose-response analysis of VSV-SARS-CoV-2 replication with fluoxetine or fluvoxamine treatment. MA104 cells were treated with compounds at 0.01 to 30 μM for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 300 μM for 25 hr. (I) Dose-response analysis of wild-type SARS-CoV-2 replication with fluoxetine or fluvoxamine treatment. Vero E6 cells were treated with compounds for 1 hr and infected with a clinical isolate of SARS-CoV-2 (MOI=0.5). S protein levels at 24 hpi were quantified based on immunofluorescence. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 300 μM for 25 hr. For all panels except A and I, experiments were repeated at least three times with similar results. Fig. 3A was performed twice. Inhibition assay in Fig. 3I was performed once and cytotoxicity assay was performed in triplicates. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (*p≤0.05; **p≤0.01; ***p≤0.001).
    Sars Cov 2 2019 Ncov Spike Rbd Antibody Rabbit Pab, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 2019 ncov spike rbd antibody rabbit pab/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike rbd antibody rabbit pab - by Bioz Stars, 2021-03
    94/100 stars
      Buy from Supplier

    94
    Sino Biological mouse igg1 fc domain
    Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human <t>IgG</t> secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.
    Mouse Igg1 Fc Domain, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse igg1 fc domain/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse igg1 fc domain - by Bioz Stars, 2021-03
    94/100 stars
      Buy from Supplier

    Image Search Results


    Viral load of the SARS‐CoV‐2‐infected rhesus macaque model. A. Average viral loads of swabs from the younger group (YG, n = 3, red line) monkeys. B. Average viral load of swabs from the elder group (EG, n = 2, blue line) monkeys. Viral loads of nasal, throat, and anal swab specimens collected from the inoculated macaques on 0, 3, 5, 7, 9, 11, and 14 dpi. C. Viral loads in varies lobe of lung tissue from YG and EG monkeys at day 7 post‐inoculation. RNA was extracted and viral load was determined by qRT‐PCR. All data are presented as mean ± SEM

    Journal: Animal Models and Experimental Medicine

    Article Title: Age‐related rhesus macaque models of COVID‐19, et al. Age‐related rhesus macaque models of COVID‐19

    doi: 10.1002/ame2.12108

    Figure Lengend Snippet: Viral load of the SARS‐CoV‐2‐infected rhesus macaque model. A. Average viral loads of swabs from the younger group (YG, n = 3, red line) monkeys. B. Average viral load of swabs from the elder group (EG, n = 2, blue line) monkeys. Viral loads of nasal, throat, and anal swab specimens collected from the inoculated macaques on 0, 3, 5, 7, 9, 11, and 14 dpi. C. Viral loads in varies lobe of lung tissue from YG and EG monkeys at day 7 post‐inoculation. RNA was extracted and viral load was determined by qRT‐PCR. All data are presented as mean ± SEM

    Article Snippet: 2.6 ELISA antibody assaySera from each animal were collected to detect the SARS‐CoV‐2 antibody through ELISA at 0, 3, 7, 11, and 14 dpi.

    Techniques: Infection, Quantitative RT-PCR

    Hematological analysis in rhesus macaques inoculated with SARS‐CoV‐2. A. The counts of white blood cells (WBC) were analysed. B. The percentage and counts of monocytes were determined. C. The percentage and counts of lymphocytes were detected. D. The percentage and counts of CD3 + CD8 + T cells, CD3 + CD4 + T cells were shown. YG (red line) and EG (blue line) were indicated in the upper right corner of each panel. All data are presented as mean ± SEM

    Journal: Animal Models and Experimental Medicine

    Article Title: Age‐related rhesus macaque models of COVID‐19, et al. Age‐related rhesus macaque models of COVID‐19

    doi: 10.1002/ame2.12108

    Figure Lengend Snippet: Hematological analysis in rhesus macaques inoculated with SARS‐CoV‐2. A. The counts of white blood cells (WBC) were analysed. B. The percentage and counts of monocytes were determined. C. The percentage and counts of lymphocytes were detected. D. The percentage and counts of CD3 + CD8 + T cells, CD3 + CD4 + T cells were shown. YG (red line) and EG (blue line) were indicated in the upper right corner of each panel. All data are presented as mean ± SEM

    Article Snippet: 2.6 ELISA antibody assaySera from each animal were collected to detect the SARS‐CoV‐2 antibody through ELISA at 0, 3, 7, 11, and 14 dpi.

    Techniques:

    The comparison of lesions in the lung between younger group (YG) and elder group (EG) by radiographic alterations, histopathological and immunohistochemical (IHC) observation of the SARS‐CoV‐2‐inoculated‐rhesus macaque. A. Anterior‐posterior thoracic X‐rays from of rhesus macaque imaged prior to SARS‐CoV‐2 inoculation (day 0) and on 7 dpi of YG and 5 dpi of EG. Areas of interstitial infiltration, indicative of pneumonia, are highlighted (red circle). Positional indicators are included (R = right). B. Histopathological changes in lungs from YG and EG. Lung tissue was collected and stained with hematoxylin and eosin. Black scale bar = 40 µm. IHC staining demonstrated that SARS‐CoV‐2 antigens were mainly in the epithelial cells and macrophages. SARS‐CoV‐2 antigens were indicated by red arrows. Red scale bar = 50 µm

    Journal: Animal Models and Experimental Medicine

    Article Title: Age‐related rhesus macaque models of COVID‐19, et al. Age‐related rhesus macaque models of COVID‐19

    doi: 10.1002/ame2.12108

    Figure Lengend Snippet: The comparison of lesions in the lung between younger group (YG) and elder group (EG) by radiographic alterations, histopathological and immunohistochemical (IHC) observation of the SARS‐CoV‐2‐inoculated‐rhesus macaque. A. Anterior‐posterior thoracic X‐rays from of rhesus macaque imaged prior to SARS‐CoV‐2 inoculation (day 0) and on 7 dpi of YG and 5 dpi of EG. Areas of interstitial infiltration, indicative of pneumonia, are highlighted (red circle). Positional indicators are included (R = right). B. Histopathological changes in lungs from YG and EG. Lung tissue was collected and stained with hematoxylin and eosin. Black scale bar = 40 µm. IHC staining demonstrated that SARS‐CoV‐2 antigens were mainly in the epithelial cells and macrophages. SARS‐CoV‐2 antigens were indicated by red arrows. Red scale bar = 50 µm

    Article Snippet: 2.6 ELISA antibody assaySera from each animal were collected to detect the SARS‐CoV‐2 antibody through ELISA at 0, 3, 7, 11, and 14 dpi.

    Techniques: Immunohistochemistry, Staining

    JIB-04 exhibits distinct post-entry antiviral mechanisms (A) Drug combination dose-response matrix and VSV-SARS-CoV-2 replication. MA104 cells were treated with JIB-04 and chloroquine or JIB-04 and NTZ for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. (B) Time of compound addition and VSV-SARS-CoV-2 replication. MA104 cells were treated with NTZ or JIB-04 (10 μM) at indicated time points relative to VSV-SARS-CoV-2 infection (MOI=3, 0 hpi). GFP signals at 8 hpi were quantified to calculate the percentage of inhibition. (C) Intracellular SARS-CoV-2 S RNA levels with JIB-04 treatment. MA104 cells were treated with JIB-04 (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1) for 1, 3, 5, and 7 hr. S RNA levels were measured by RT-qPCR. (D) Western blot analysis of SARS-CoV-2 S protein levels with JIB-04 treatment. MA104 cells were treated with JIB-04 (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1) for 1, 3, 5, and 7 hr. FL: full-length. S2: cleaved S2 fragment. (* non-specific band) (E) Histone demethylase siRNA knockdown and RV replication. HEK293 cells were transfected with scrambled siRNA or siRNA targeting indicated histone demethylases for 48 hr and infected with porcine RV (MOI=0.01). Viral RNA copy numbers at 12 hpi were quantified by RT-qPCR. (F) Volcano plot of differentially expressed transcripts with JIB-04 treatment and RV infection. HEK293 cells were treated with DMSO or JIB-04 (10 μM) for 12 hr, and mock-infected (left panel) or infected with porcine RV (MOI=0.01, right panel) for another 12 hr. Red dots represent upregulated genes and green dots represent downregulated genes in JIB-04 treated cells. (G) Expression of three top genes in (F) with JIB-04 treatment. HEK293 cells were treated with JIB-04 (10 μM) for 12 hr and mock-infected or infected porcine RV (MOI=0.01) for 12 hr. mRNA levels of CYP1A1, CYP1B1, and AHRR at 12 hpi were measured by RT-qPCR. (H) Dose-response analysis of VSV-SARS-CoV-2 replication with fluoxetine or fluvoxamine treatment. MA104 cells were treated with compounds at 0.01 to 30 μM for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 300 μM for 25 hr. (I) Dose-response analysis of wild-type SARS-CoV-2 replication with fluoxetine or fluvoxamine treatment. Vero E6 cells were treated with compounds for 1 hr and infected with a clinical isolate of SARS-CoV-2 (MOI=0.5). S protein levels at 24 hpi were quantified based on immunofluorescence. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 300 μM for 25 hr. For all panels except A and I, experiments were repeated at least three times with similar results. Fig. 3A was performed twice. Inhibition assay in Fig. 3I was performed once and cytotoxicity assay was performed in triplicates. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (*p≤0.05; **p≤0.01; ***p≤0.001).

    Journal: bioRxiv

    Article Title: Nitazoxanide and JIB-04 have broad-spectrum antiviral activity and inhibit SARS-CoV-2 replication in cell culture and coronavirus pathogenesis in a pig model

    doi: 10.1101/2020.09.24.312165

    Figure Lengend Snippet: JIB-04 exhibits distinct post-entry antiviral mechanisms (A) Drug combination dose-response matrix and VSV-SARS-CoV-2 replication. MA104 cells were treated with JIB-04 and chloroquine or JIB-04 and NTZ for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. (B) Time of compound addition and VSV-SARS-CoV-2 replication. MA104 cells were treated with NTZ or JIB-04 (10 μM) at indicated time points relative to VSV-SARS-CoV-2 infection (MOI=3, 0 hpi). GFP signals at 8 hpi were quantified to calculate the percentage of inhibition. (C) Intracellular SARS-CoV-2 S RNA levels with JIB-04 treatment. MA104 cells were treated with JIB-04 (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1) for 1, 3, 5, and 7 hr. S RNA levels were measured by RT-qPCR. (D) Western blot analysis of SARS-CoV-2 S protein levels with JIB-04 treatment. MA104 cells were treated with JIB-04 (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1) for 1, 3, 5, and 7 hr. FL: full-length. S2: cleaved S2 fragment. (* non-specific band) (E) Histone demethylase siRNA knockdown and RV replication. HEK293 cells were transfected with scrambled siRNA or siRNA targeting indicated histone demethylases for 48 hr and infected with porcine RV (MOI=0.01). Viral RNA copy numbers at 12 hpi were quantified by RT-qPCR. (F) Volcano plot of differentially expressed transcripts with JIB-04 treatment and RV infection. HEK293 cells were treated with DMSO or JIB-04 (10 μM) for 12 hr, and mock-infected (left panel) or infected with porcine RV (MOI=0.01, right panel) for another 12 hr. Red dots represent upregulated genes and green dots represent downregulated genes in JIB-04 treated cells. (G) Expression of three top genes in (F) with JIB-04 treatment. HEK293 cells were treated with JIB-04 (10 μM) for 12 hr and mock-infected or infected porcine RV (MOI=0.01) for 12 hr. mRNA levels of CYP1A1, CYP1B1, and AHRR at 12 hpi were measured by RT-qPCR. (H) Dose-response analysis of VSV-SARS-CoV-2 replication with fluoxetine or fluvoxamine treatment. MA104 cells were treated with compounds at 0.01 to 30 μM for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 300 μM for 25 hr. (I) Dose-response analysis of wild-type SARS-CoV-2 replication with fluoxetine or fluvoxamine treatment. Vero E6 cells were treated with compounds for 1 hr and infected with a clinical isolate of SARS-CoV-2 (MOI=0.5). S protein levels at 24 hpi were quantified based on immunofluorescence. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 300 μM for 25 hr. For all panels except A and I, experiments were repeated at least three times with similar results. Fig. 3A was performed twice. Inhibition assay in Fig. 3I was performed once and cytotoxicity assay was performed in triplicates. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (*p≤0.05; **p≤0.01; ***p≤0.001).

    Article Snippet: Proteins were resolved in SDS-PAGE and detected as describedusing the following antibodies: GAPDH (631402, Biolegend), rotavirus VP6 (rabbit polyclonal, ABclonal technology), and SARS-CoV-2 S2 (40592-T62, Sino Biological).

    Techniques: Infection, Inhibition, Quantitative RT-PCR, Western Blot, Transfection, Expressing, Immunofluorescence, Cytotoxicity Assay

    Nitazoxanide and JIB-04 inhibit SARS-CoV-2 replication (A) Chemical structures of NTZ and JIB-04 E-isomer from ChemSpider database. (B) Representative images of Vero E6 cells infected by SARS-CoV-2-mNeonGreen (MOI=0.5) at 24 hpi in Fig. 1A . Experiments were repeated at least three times with similar results.

    Journal: bioRxiv

    Article Title: Nitazoxanide and JIB-04 have broad-spectrum antiviral activity and inhibit SARS-CoV-2 replication in cell culture and coronavirus pathogenesis in a pig model

    doi: 10.1101/2020.09.24.312165

    Figure Lengend Snippet: Nitazoxanide and JIB-04 inhibit SARS-CoV-2 replication (A) Chemical structures of NTZ and JIB-04 E-isomer from ChemSpider database. (B) Representative images of Vero E6 cells infected by SARS-CoV-2-mNeonGreen (MOI=0.5) at 24 hpi in Fig. 1A . Experiments were repeated at least three times with similar results.

    Article Snippet: Proteins were resolved in SDS-PAGE and detected as describedusing the following antibodies: GAPDH (631402, Biolegend), rotavirus VP6 (rabbit polyclonal, ABclonal technology), and SARS-CoV-2 S2 (40592-T62, Sino Biological).

    Techniques: Infection

    Nitazoxanide and JIB-04 inhibit the replication of multiple viruses (A) Mean fluorescence intensity of GFP positive cells in Fig. 2B was quantified by flow cytometry. (B) Dose-response analysis of VSV-SARS-CoV-2 replication with NTZ or JIB-04 treatment. MA104 cells were treated with compounds at indicated concentrations for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). At 24 hpi, images of GFP positive infected cells were acquired by the ECHO fluorescence microscope. (C) Same as (B) except that cells were infected with an MOI of 0.1. (D) Dose-response analysis of intracellular viral RNA levels with NTZ, JIB-04, or chloroquine treatment. MA104 cells were treated with compounds at 0.1 to 30 μM for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). VSV RNA levels at 24 hpi were measured by RT-qPCR. (E) Western blot analysis of RV antigen VP6 levels with JIB-04 treatment. HEK293 cells were treated with JIB-04 at 1, 5, or 10 μM for 6 hr and infected with porcine RV (MOI=0.01) for 12 hr. GAPDH was used as a loading control. All experiments were repeated at least three times with similar results. Data are represented as mean ± SEM.

    Journal: bioRxiv

    Article Title: Nitazoxanide and JIB-04 have broad-spectrum antiviral activity and inhibit SARS-CoV-2 replication in cell culture and coronavirus pathogenesis in a pig model

    doi: 10.1101/2020.09.24.312165

    Figure Lengend Snippet: Nitazoxanide and JIB-04 inhibit the replication of multiple viruses (A) Mean fluorescence intensity of GFP positive cells in Fig. 2B was quantified by flow cytometry. (B) Dose-response analysis of VSV-SARS-CoV-2 replication with NTZ or JIB-04 treatment. MA104 cells were treated with compounds at indicated concentrations for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). At 24 hpi, images of GFP positive infected cells were acquired by the ECHO fluorescence microscope. (C) Same as (B) except that cells were infected with an MOI of 0.1. (D) Dose-response analysis of intracellular viral RNA levels with NTZ, JIB-04, or chloroquine treatment. MA104 cells were treated with compounds at 0.1 to 30 μM for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3). VSV RNA levels at 24 hpi were measured by RT-qPCR. (E) Western blot analysis of RV antigen VP6 levels with JIB-04 treatment. HEK293 cells were treated with JIB-04 at 1, 5, or 10 μM for 6 hr and infected with porcine RV (MOI=0.01) for 12 hr. GAPDH was used as a loading control. All experiments were repeated at least three times with similar results. Data are represented as mean ± SEM.

    Article Snippet: Proteins were resolved in SDS-PAGE and detected as describedusing the following antibodies: GAPDH (631402, Biolegend), rotavirus VP6 (rabbit polyclonal, ABclonal technology), and SARS-CoV-2 S2 (40592-T62, Sino Biological).

    Techniques: Fluorescence, Flow Cytometry, Infection, Microscopy, Quantitative RT-PCR, Western Blot

    Nitazoxanide and JIB-04 broadly inhibit DNA and RNA viruses in different cell types (A) Dose-response analysis of VSV and VSV-SARS-CoV-2 replication with 15 compounds. MA104 cells were treated with indicated compounds at 0.01 to 30 μM for 1 hr and infected with VSV or VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. EC 50 values for VSV and VSV-SARS-CoV-2 are shown in each graph in red and blue, respectively. (B) Virus infectivity with NTZ or JIB-04 treatment. Vero E6-TMPRSS2 cells were treated with compounds (10 μM) for 1 hr and infected with VSV or VSV-SARS-CoV-2 (MOI=3). At 6 hpi, percentages of GFP positive cells were quantified by flow cytometry. (C) Dose-response analysis of VSV-SARS-CoV-2 replication and cytotoxicity with NTZ or JIB-04 treatment. For EC 50 measurement, MA104 cells were treated with compounds at 0.01 to 30 μM for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3) for 24 hr. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 3 mM for 25 hr. SI: selectivity index. (D) Intracellular viral RNA levels with NTZ or JIB-04 treatment. MA104 cells were treated with compounds (10 μM) for 1 hr and infected with vaccinia virus (VACV), herpes simplex virus-1 (HSV-1), or rotavirus (RV, RRV and UK strains) (MOI=1). Viral RNA levels at 24 hpi were measured by RT-qPCR for VACV B10R, HSV-1 ICP-27, and RV NSP5, respectively. (E) Viral RNA copy numbers with JIB-04 treatment. HEK293 cells were treated with JIB-04 (10 μM) for 6 hr and infected with porcine rotavirus (MOI=0.01) for 6 hr. ST cells were treated with JIB-04 (10 μM) for 12 hr and infected with transmissible gastroenteritis virus (TGEV) (MOI=0.01) for 12 hr. Viral RNA copy numbers were measured by RT-qPCR. (F) TGEV titers in the cell supernatant with JIB-04 treatment. ST cells were treated with JIB-04 (10 μM) for 12 hr and infected with TGEV (MOI=0.01). Virus titers at 6 and 12 hpi were measured by plaque assays. (G) Intracellular viral RNA levels with NTZ or JIB-04 treatment in different cell types. HEK293-hACE2, HEK293-hACE2-TMPRSS2, and Calu-3 cells were treated with compounds (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1). VSV RNA levels at 24 hpi were measured by RT-qPCR. For all panels except A, experiments were repeated at least three times with similar results. Fig. 2A was performed once. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (*p≤0.05; **p≤0.01; ***p≤0.001).

    Journal: bioRxiv

    Article Title: Nitazoxanide and JIB-04 have broad-spectrum antiviral activity and inhibit SARS-CoV-2 replication in cell culture and coronavirus pathogenesis in a pig model

    doi: 10.1101/2020.09.24.312165

    Figure Lengend Snippet: Nitazoxanide and JIB-04 broadly inhibit DNA and RNA viruses in different cell types (A) Dose-response analysis of VSV and VSV-SARS-CoV-2 replication with 15 compounds. MA104 cells were treated with indicated compounds at 0.01 to 30 μM for 1 hr and infected with VSV or VSV-SARS-CoV-2 (MOI=3). GFP signals at 24 hpi were quantified to calculate the percentage of inhibition. EC 50 values for VSV and VSV-SARS-CoV-2 are shown in each graph in red and blue, respectively. (B) Virus infectivity with NTZ or JIB-04 treatment. Vero E6-TMPRSS2 cells were treated with compounds (10 μM) for 1 hr and infected with VSV or VSV-SARS-CoV-2 (MOI=3). At 6 hpi, percentages of GFP positive cells were quantified by flow cytometry. (C) Dose-response analysis of VSV-SARS-CoV-2 replication and cytotoxicity with NTZ or JIB-04 treatment. For EC 50 measurement, MA104 cells were treated with compounds at 0.01 to 30 μM for 1 hr and infected with VSV-SARS-CoV-2 (MOI=3) for 24 hr. For CC 50 measurement, cells were treated with compounds at 0.1 μM to 3 mM for 25 hr. SI: selectivity index. (D) Intracellular viral RNA levels with NTZ or JIB-04 treatment. MA104 cells were treated with compounds (10 μM) for 1 hr and infected with vaccinia virus (VACV), herpes simplex virus-1 (HSV-1), or rotavirus (RV, RRV and UK strains) (MOI=1). Viral RNA levels at 24 hpi were measured by RT-qPCR for VACV B10R, HSV-1 ICP-27, and RV NSP5, respectively. (E) Viral RNA copy numbers with JIB-04 treatment. HEK293 cells were treated with JIB-04 (10 μM) for 6 hr and infected with porcine rotavirus (MOI=0.01) for 6 hr. ST cells were treated with JIB-04 (10 μM) for 12 hr and infected with transmissible gastroenteritis virus (TGEV) (MOI=0.01) for 12 hr. Viral RNA copy numbers were measured by RT-qPCR. (F) TGEV titers in the cell supernatant with JIB-04 treatment. ST cells were treated with JIB-04 (10 μM) for 12 hr and infected with TGEV (MOI=0.01). Virus titers at 6 and 12 hpi were measured by plaque assays. (G) Intracellular viral RNA levels with NTZ or JIB-04 treatment in different cell types. HEK293-hACE2, HEK293-hACE2-TMPRSS2, and Calu-3 cells were treated with compounds (10 μM) for 1 hr and infected with VSV-SARS-CoV-2 (MOI=1). VSV RNA levels at 24 hpi were measured by RT-qPCR. For all panels except A, experiments were repeated at least three times with similar results. Fig. 2A was performed once. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (*p≤0.05; **p≤0.01; ***p≤0.001).

    Article Snippet: Proteins were resolved in SDS-PAGE and detected as describedusing the following antibodies: GAPDH (631402, Biolegend), rotavirus VP6 (rabbit polyclonal, ABclonal technology), and SARS-CoV-2 S2 (40592-T62, Sino Biological).

    Techniques: Infection, Inhibition, Flow Cytometry, Quantitative RT-PCR

    Nitazoxanide and JIB-04 inhibit SARS-CoV-2 replication (A) Small molecule inhibitor screen. Vero E6 cells were treated with individual compounds (listed in Table S1) at 10 μM for 1 hour (hr) and infected with SARS-CoV-2-mNeonGreen (MOI=0.5). At 24 hr post infection (hpi), cells were fixed and nuclei were stained by Hoechst 33342. The intensities of mNeonGreen and Hoechst were quantified by the Typhoon biomolecular imager and Cytation plate reader, respectively. The ratio of mNeonGreen and Hoechst is plotted as percentage of inhibition. (B) Dose-response analysis of wild-type SARS-CoV-2 replication with NTZ or JIB-04 treatment. Vero E6 cells were treated with compounds for 1 hr and infected with a clinical isolate of SARS-CoV-2 (MOI=0.5). S protein levels were quantified at 24 hpi based on immunofluorescence. For CC 50 measurement, cells were treated with inhibitors at 0.3 μM to 1 mM for 25 hr. SI: selectivity index. (C) Dose-response analysis of intracellular viral RNA levels with compounds. Vero E6 cells were treated with NTZ (10 μM), JIB-04 (10 μM), chloroquine (10 μM), remdesivir (3 μM), or camostat (10 μM) for 1 hr and infected with a clinical isolate of SARS-CoV-2 (MOI=0.5). SARS-CoV-2 RNA levels at 24 hpi were measured by RT-qPCR. For all panels except A, experiments were repeated at least three times with similar results. Fig. 1A was performed once with raw data included in Dataset S1. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (*p≤0.05).

    Journal: bioRxiv

    Article Title: Nitazoxanide and JIB-04 have broad-spectrum antiviral activity and inhibit SARS-CoV-2 replication in cell culture and coronavirus pathogenesis in a pig model

    doi: 10.1101/2020.09.24.312165

    Figure Lengend Snippet: Nitazoxanide and JIB-04 inhibit SARS-CoV-2 replication (A) Small molecule inhibitor screen. Vero E6 cells were treated with individual compounds (listed in Table S1) at 10 μM for 1 hour (hr) and infected with SARS-CoV-2-mNeonGreen (MOI=0.5). At 24 hr post infection (hpi), cells were fixed and nuclei were stained by Hoechst 33342. The intensities of mNeonGreen and Hoechst were quantified by the Typhoon biomolecular imager and Cytation plate reader, respectively. The ratio of mNeonGreen and Hoechst is plotted as percentage of inhibition. (B) Dose-response analysis of wild-type SARS-CoV-2 replication with NTZ or JIB-04 treatment. Vero E6 cells were treated with compounds for 1 hr and infected with a clinical isolate of SARS-CoV-2 (MOI=0.5). S protein levels were quantified at 24 hpi based on immunofluorescence. For CC 50 measurement, cells were treated with inhibitors at 0.3 μM to 1 mM for 25 hr. SI: selectivity index. (C) Dose-response analysis of intracellular viral RNA levels with compounds. Vero E6 cells were treated with NTZ (10 μM), JIB-04 (10 μM), chloroquine (10 μM), remdesivir (3 μM), or camostat (10 μM) for 1 hr and infected with a clinical isolate of SARS-CoV-2 (MOI=0.5). SARS-CoV-2 RNA levels at 24 hpi were measured by RT-qPCR. For all panels except A, experiments were repeated at least three times with similar results. Fig. 1A was performed once with raw data included in Dataset S1. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (*p≤0.05).

    Article Snippet: Proteins were resolved in SDS-PAGE and detected as describedusing the following antibodies: GAPDH (631402, Biolegend), rotavirus VP6 (rabbit polyclonal, ABclonal technology), and SARS-CoV-2 S2 (40592-T62, Sino Biological).

    Techniques: Infection, Staining, Inhibition, Immunofluorescence, Quantitative RT-PCR

    Inhibition or knockdown of specific KDM histone demethylases inhibits virus replication (A) Expression of IFN and IFN-stimulated genes with NTZ or JIB-04 treatment. HEK293 cells were treated with NTZ (10 μM), JIB-04 (3 μM), or transfected with low-molecular-weight poly(I:C) (100 ng/ml) for 24 hr. mRNA levels of IFNL3 and CXCL10 were measured by RT-qPCR. (B) Autophagy formation with compound treatment. HEK293 cells were transfected with EGFP-LC3 plasmid for 24 hr and treated with rapamycin (100 nM), NTZ (10 μM), or JIB-04 (3 μM) for another 18 hr. GFP positive punctate structures indicate autophagy activation. Scale bar, 20 μm. (C) Intracellular viral RNA levels with JIB-04 and camostat treatment. Calu-3 cells were treated with compounds (10 μM) for 1 hr and infected with VSV-SARS-CoV (MOI=3). VSV RNA levels at 24 hpi were measured by RT-qPCR. (D) Intracellular SARS-CoV-2 S RNA levels with JIB-04 treatment. HEK293 cells were transfected with SARS-CoV-2 S plasmid for 2 hr and treated with JIB-04 (3 μM) for 24 hr. S RNA levels were measured by RT-qPCR. (E) Western blot analysis of SARS-CoV-2 S protein levels with JIB-04 treatment. HEK293 cells were transfected with SARS-CoV-2 S plasmid for 2 hr and treated with JIB-04 (3 μM) for 24 hr. FL: full-length. S2: cleaved S2 fragment. (F) siRNA-mediated knockdown of JIB-04 target histone demethylases. HEK293 cells were transfected with scrambled siRNA or siRNA targeting indicated histone demethylases for 48 hr. mRNA levels of indicated histone demethylases were measured by RT-qPCR. (G) Western blot analysis of RV antigen VP6 levels in cells with histone demethylase siRNA knockdown. HEK293 cells were transfected with scrambled siRNA or siRNA targeting indicated histone demethylases for 48 hr and infected with porcine RV (MOI=0.01) for 12 hr. (H) Pathway enrichment analysis of gene expression regulated by JIB-04 treatment. Downregulated genes in Fig. 3F with p values

    Journal: bioRxiv

    Article Title: Nitazoxanide and JIB-04 have broad-spectrum antiviral activity and inhibit SARS-CoV-2 replication in cell culture and coronavirus pathogenesis in a pig model

    doi: 10.1101/2020.09.24.312165

    Figure Lengend Snippet: Inhibition or knockdown of specific KDM histone demethylases inhibits virus replication (A) Expression of IFN and IFN-stimulated genes with NTZ or JIB-04 treatment. HEK293 cells were treated with NTZ (10 μM), JIB-04 (3 μM), or transfected with low-molecular-weight poly(I:C) (100 ng/ml) for 24 hr. mRNA levels of IFNL3 and CXCL10 were measured by RT-qPCR. (B) Autophagy formation with compound treatment. HEK293 cells were transfected with EGFP-LC3 plasmid for 24 hr and treated with rapamycin (100 nM), NTZ (10 μM), or JIB-04 (3 μM) for another 18 hr. GFP positive punctate structures indicate autophagy activation. Scale bar, 20 μm. (C) Intracellular viral RNA levels with JIB-04 and camostat treatment. Calu-3 cells were treated with compounds (10 μM) for 1 hr and infected with VSV-SARS-CoV (MOI=3). VSV RNA levels at 24 hpi were measured by RT-qPCR. (D) Intracellular SARS-CoV-2 S RNA levels with JIB-04 treatment. HEK293 cells were transfected with SARS-CoV-2 S plasmid for 2 hr and treated with JIB-04 (3 μM) for 24 hr. S RNA levels were measured by RT-qPCR. (E) Western blot analysis of SARS-CoV-2 S protein levels with JIB-04 treatment. HEK293 cells were transfected with SARS-CoV-2 S plasmid for 2 hr and treated with JIB-04 (3 μM) for 24 hr. FL: full-length. S2: cleaved S2 fragment. (F) siRNA-mediated knockdown of JIB-04 target histone demethylases. HEK293 cells were transfected with scrambled siRNA or siRNA targeting indicated histone demethylases for 48 hr. mRNA levels of indicated histone demethylases were measured by RT-qPCR. (G) Western blot analysis of RV antigen VP6 levels in cells with histone demethylase siRNA knockdown. HEK293 cells were transfected with scrambled siRNA or siRNA targeting indicated histone demethylases for 48 hr and infected with porcine RV (MOI=0.01) for 12 hr. (H) Pathway enrichment analysis of gene expression regulated by JIB-04 treatment. Downregulated genes in Fig. 3F with p values

    Article Snippet: Proteins were resolved in SDS-PAGE and detected as describedusing the following antibodies: GAPDH (631402, Biolegend), rotavirus VP6 (rabbit polyclonal, ABclonal technology), and SARS-CoV-2 S2 (40592-T62, Sino Biological).

    Techniques: Inhibition, Expressing, Transfection, Molecular Weight, Quantitative RT-PCR, Plasmid Preparation, Activation Assay, Infection, Western Blot

    Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human IgG secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.

    Journal: bioRxiv

    Article Title: Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein

    doi: 10.1101/2020.05.12.091462

    Figure Lengend Snippet: Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human IgG secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.

    Article Snippet: RBD protein fused to mouse IgG1 Fc domain (designated RBD-mFc), was purchased from Sino Biological (40592-V05H).

    Techniques: Functional Assay, Binding Assay, Fluorescence, Clone Assay

    Changes of laboratory results before and at day 1–5 after convalescent plasma transfusion. a , b SARS-CoV-2 specific IgG and IgM levels, respectively, determined by MCLIA. c , d Cycle threshold (Ct) values of ORF1ab-gene and N-gene, respectively. A Ct value of 40 was defined as undetectable. e PaO 2 /FiO 2 (normal range: 400–500 mmHg). f White blood cell count (normal range: 3.5–9.5). g Lymphocyte count (normal range: 1.1–3.2). h C-reactive protein (normal range:

    Journal: Signal Transduction and Targeted Therapy

    Article Title: The efficacy assessment of convalescent plasma therapy for COVID-19 patients: a multi-center case series

    doi: 10.1038/s41392-020-00329-x

    Figure Lengend Snippet: Changes of laboratory results before and at day 1–5 after convalescent plasma transfusion. a , b SARS-CoV-2 specific IgG and IgM levels, respectively, determined by MCLIA. c , d Cycle threshold (Ct) values of ORF1ab-gene and N-gene, respectively. A Ct value of 40 was defined as undetectable. e PaO 2 /FiO 2 (normal range: 400–500 mmHg). f White blood cell count (normal range: 3.5–9.5). g Lymphocyte count (normal range: 1.1–3.2). h C-reactive protein (normal range:

    Article Snippet: 80 ng/ml recombinant SARS-CoV-2 Spike RBD-mFc (Sino Biological) was mixed with the presence or absence of serially diluted CP or serum samples 1:1 and incubated at 37 °C for 1 h, then add the 100 μl mixed solution to the wells.

    Techniques: Cell Counting

    SARS-CoV-2 specific antibody levels of CP samples measured by serology tests, receptor-binding assay, and pseudovirus based neutralization assay. a The correlations among anti-SARS-CoV-2 specific IgG and IgM titers detected by commercial MCLIA kits, anti-S-RBD and anti-NP specific IgG titers determined by in-house ELISA assays, inhibition activity measured by a receptor-binding assay, and neutralizing antibody titer measured by a pseudovirus based neutralization assay. b Comparisons of antibody levels between CP samples collected before and after 21 days from symptom onset. MCLIA magnetic chemiluminescence enzyme immunoassay, ELISA enzyme-linked immunosorbent assay, RBD receptor binding domains, NP nucleoprotein, IT50 inhibitory titer which was calculated with the dilution of plasma that inhibits 50% RBD-Fc binding to receptor ACE2, NAT50 neutralizing antibody titer which was calculated with the highest dilution of plasma that resulted in a 50% reduction of virus infection, GMT geometric mean titer, CI confidence interval

    Journal: Signal Transduction and Targeted Therapy

    Article Title: The efficacy assessment of convalescent plasma therapy for COVID-19 patients: a multi-center case series

    doi: 10.1038/s41392-020-00329-x

    Figure Lengend Snippet: SARS-CoV-2 specific antibody levels of CP samples measured by serology tests, receptor-binding assay, and pseudovirus based neutralization assay. a The correlations among anti-SARS-CoV-2 specific IgG and IgM titers detected by commercial MCLIA kits, anti-S-RBD and anti-NP specific IgG titers determined by in-house ELISA assays, inhibition activity measured by a receptor-binding assay, and neutralizing antibody titer measured by a pseudovirus based neutralization assay. b Comparisons of antibody levels between CP samples collected before and after 21 days from symptom onset. MCLIA magnetic chemiluminescence enzyme immunoassay, ELISA enzyme-linked immunosorbent assay, RBD receptor binding domains, NP nucleoprotein, IT50 inhibitory titer which was calculated with the dilution of plasma that inhibits 50% RBD-Fc binding to receptor ACE2, NAT50 neutralizing antibody titer which was calculated with the highest dilution of plasma that resulted in a 50% reduction of virus infection, GMT geometric mean titer, CI confidence interval

    Article Snippet: 80 ng/ml recombinant SARS-CoV-2 Spike RBD-mFc (Sino Biological) was mixed with the presence or absence of serially diluted CP or serum samples 1:1 and incubated at 37 °C for 1 h, then add the 100 μl mixed solution to the wells.

    Techniques: Reporter Assay, Neutralization, Enzyme-linked Immunosorbent Assay, Inhibition, Activity Assay, Binding Assay, Infection