anti spike rbd antibodies  (Sino Biological)


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    Name:
    MERS CoV Spike Protein Antibody Rabbit PAb
    Description:
    Produced in rabbits immunized with purified recombinant MERS CoV NCoV Novel coronavirus Spike Protein ECD aa 1 1297 Catalog 40069 V08B AFS88936 1 Met1 Trp1297 MERS CoV NCoV Novel coronavirus Spike Protein ECD aa 1 1297 specific IgG was purified by MERS CoV NCoV Novel coronavirus Spike Protein ECD aa 1 1297 affinity chromatography
    Catalog Number:
    40069-T30
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    MERS CoV
    Applications:
    ELISA
    Immunogen:
    Recombinant MERS-CoV (NCoV / Novel coronavirus) Spike Protein (ECD, aa 1-1297) Protein (Catalog#40069-V08B)
    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-RBD Antibody, Anti-S Antibody, Anti-s1 Antibody, Anti-Spike RBD Antibody
    Antibody Type:
    PAb
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Sino Biological anti spike rbd antibodies
    Produced in rabbits immunized with purified recombinant MERS CoV NCoV Novel coronavirus Spike Protein ECD aa 1 1297 Catalog 40069 V08B AFS88936 1 Met1 Trp1297 MERS CoV NCoV Novel coronavirus Spike Protein ECD aa 1 1297 specific IgG was purified by MERS CoV NCoV Novel coronavirus Spike Protein ECD aa 1 1297 affinity chromatography
    https://www.bioz.com/result/anti spike rbd antibodies/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti spike rbd antibodies - by Bioz Stars, 2021-06
    95/100 stars

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    Related Articles

    Incubation:

    Article Title: Characterization of a human monoclonal antibody generated from a B-cell specific for a prefusion-stabilized spike protein of Middle East respiratory syndrome coronavirus
    Article Snippet: Then, human anti-MERS-CoV antibodies were incubated with the Vero cells infected with MERS-CoV. .. As positive controls, a rabbit polyclonal anti-MERS-CoV spike protein antibody (Sino biological Inc., Beijing, China) for MERS-CoV, a mouse anti-229E coronavirus nucleoprotein OC-43 antibody (MERCK, Darmstadt, Germany) for HCoV-229E, a mouse anti-coronavirus antibody, hCoV OC-43 (LifeSpan BioScience, Seattle, WA, USA) for HCoV-OC43, a rabbit polyclonal anti-hCoV-HKU1 spike protein antibody (Sino Biological Inc., Beijing, China) for hCoV-NL63, a human serum of a Korean SARS-CoV-2 convalescent person were also incubated. .. The secondary antibodies, Fluorescein isothiocyanate (FITC) -conjugated goat anti-human-IgG (Abcam, Cambridge, U.K.), FITC-conjugated goat anti-rabbit-IgG (Abcam, Cambridge, U.K.), and FITC-conjugated goat anti-mouse-IgG (Jackson Immunoresearch, West Grove, PA, USA) were added to the cells and incubated.

    Article Title: A Novel Bacterium-Like Particle Vaccine Displaying the MERS-CoV Receptor-Binding Domain Induces Specific Mucosal and Systemic Immune Responses in Mice
    Article Snippet: Briefly, Sf9 cells cultured in 96-well plates at 2 × 106 cells/mL were infected with the recombinant baculovirus. .. After 48 h of infection, the cultured plates were fixed with 80% cold acetone overnight at -20°C, washed three times with PBS-0.05% Tween 20 (PBST), and then incubated with a rabbit anti-MERS-S polyclonal antibody (1:500, Sino Biological Inc, Beijing, China) containing 1% bovine serum albumin (BSA, Sigma-Aldrich, USA) at 37°C for 1 h. After three washes with PBST, an FITC-labeled goat against rabbit IgG antibody (1:300, BioWorld, Inc, St. Louis, MN, USA) was added with Evans blue (Sigma-Aldrich, St. Louis, MN, USA) for 1 h at 37 °C. ..

    Article Title: A high-throughput inhibition assay to study MERS-CoV antibody interactions using image cytometry
    Article Snippet: After incubation, cells were washed with PBS, fixed with 80% cold acetone for 20 min at RT, and washed again with PBS. .. Subsequently, 100 μL of MERS-CoV S rabbit polyclonal antibodies (Sino Biological, Beijing, China) was pipetted into each well, incubated for 60 min at RT, and washed twice. .. Next, secondary goat anti-rabbit IgG H & L labeled with Alexa Fluor® 488 (AF488) and DPP4 PE-conjugated mouse antibody (Sino Biological) were added, incubated for 60 min at RT, and washed twice.

    Infection:

    Article Title: A Novel Bacterium-Like Particle Vaccine Displaying the MERS-CoV Receptor-Binding Domain Induces Specific Mucosal and Systemic Immune Responses in Mice
    Article Snippet: Briefly, Sf9 cells cultured in 96-well plates at 2 × 106 cells/mL were infected with the recombinant baculovirus. .. After 48 h of infection, the cultured plates were fixed with 80% cold acetone overnight at -20°C, washed three times with PBS-0.05% Tween 20 (PBST), and then incubated with a rabbit anti-MERS-S polyclonal antibody (1:500, Sino Biological Inc, Beijing, China) containing 1% bovine serum albumin (BSA, Sigma-Aldrich, USA) at 37°C for 1 h. After three washes with PBST, an FITC-labeled goat against rabbit IgG antibody (1:300, BioWorld, Inc, St. Louis, MN, USA) was added with Evans blue (Sigma-Aldrich, St. Louis, MN, USA) for 1 h at 37 °C. ..

    Cell Culture:

    Article Title: A Novel Bacterium-Like Particle Vaccine Displaying the MERS-CoV Receptor-Binding Domain Induces Specific Mucosal and Systemic Immune Responses in Mice
    Article Snippet: Briefly, Sf9 cells cultured in 96-well plates at 2 × 106 cells/mL were infected with the recombinant baculovirus. .. After 48 h of infection, the cultured plates were fixed with 80% cold acetone overnight at -20°C, washed three times with PBS-0.05% Tween 20 (PBST), and then incubated with a rabbit anti-MERS-S polyclonal antibody (1:500, Sino Biological Inc, Beijing, China) containing 1% bovine serum albumin (BSA, Sigma-Aldrich, USA) at 37°C for 1 h. After three washes with PBST, an FITC-labeled goat against rabbit IgG antibody (1:300, BioWorld, Inc, St. Louis, MN, USA) was added with Evans blue (Sigma-Aldrich, St. Louis, MN, USA) for 1 h at 37 °C. ..

    Immunohistochemistry:

    Article Title: Intratracheal exposure of common marmosets to MERS-CoV Jordan-n3/2012 or MERS-CoV EMC/2012 isolates does not result in lethal disease
    Article Snippet: .. To detect MERS-CoV antigen, IHC was performed using a rabbit polyclonal antiserum against MERS-CoV (Sino Biological PR China) (1:1000). .. Tissues from an uninfected control animal were used to validate all IHC procedures.

    Recombinant:

    Article Title: Development of a SERS-based lateral flow immunoassay for rapid and ultra-sensitive detection of anti-SARS-CoV-2 IgM/IgG in clinical samples
    Article Snippet: 2.1 Materials and chemicals Tetraethyl orthosilicate (TEOS), 2-(N-morpholino) ethanesulfonic (MES), polyethyleneimine (PEI, 25 K), polyvinylpyrrolidone (PVP, 40 K), Tween 20, N-hydroxysuccinimide (NHS), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB), sodium borohydride (NaBH4 ), trisodium citrate (Na3 C6 H5 O7 , TSC), goat anti-human IgM, goat anti-human IgG, goat anti-rabbit IgG, bovine serum albumin (BSA) were obtained from Sigma-Aldrich (USA). .. SARS-CoV-2 S protein (recombinant) and S protein antibody (Ab) were obtained from Sino Biological Inc. (Beijing, China). .. Chloroauric acid tetrahydrate, sodium borohydride, formaldehyde (37%, w/w), ammonia solution (28%, w/w), chloroauric acid tetrahydrate (HAuCl4 ·4H2 O), and silver nitrate (AgNO3) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).

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  • 94
    Sino Biological sars cov 2 rbd
    RU169 output clone diversity Using the <t>SARS-CoV-2</t> RBD as the target of library panning and FACS selection for screen RU169 produced a high number of unique clones, indicating high, unexplored, diversity in the output.
    Sars Cov 2 Rbd, 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 rbd/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 rbd - by Bioz Stars, 2021-06
    94/100 stars
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    95
    Sino Biological sars cov 2 2019 ncov spike rbd antibody rabbit pab
    ELISA ( x -axis) vs. LFRET ( y -axis) results by disease severity. ( a ) Anti-NP IgA ELISA vs. anti-NP LFRET (N = 81, R = 0.25). ( b ) anti-NP IgG ELISA vs. anti-NP LFRET (N = 129, R = 0.62). ( c ) anti-NP IgM ELISA vs. anti-NP LFRET (N = 81, R = 0.13). ( d ) anti-SP IgA ELISA vs. anti-SP LFRET (N = 129, R = 0.53). ( e ) anti-SP IgG ELISA vs. anti-SP LFRET (N = 129, R = 0.62). ( f ) anti-SP IgM ELISA vs. anti-SP LFRET (N = 81, R = 0.56). Color of the dot indicates <t>SARS-CoV-2</t> PCR result and disease severity: cyan = PCR negative; yellow = non-hospitalized, PCR-positive; red = non-ICU hospitalized, PCR positive; black = hospitalized in ICU, PCR positive. Horizontal and vertical black lines indicate LFRET and ELISA cutoffs. On the x -axis, ELISA absorbance on a logarithmic scale and on the y -axis, LFRET signal on a logarithmic scale. SP = spike glycoprotein. NP = nucleoprotein. LFRET = protein L–based time-resolved Förster resonance energy transfer immunoassay. ELISA = enzyme immunoassay. R = Pearson’s correlation coefficient.
    Sars Cov 2 2019 Ncov Spike Rbd Antibody Rabbit Pab, 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 2019 ncov spike rbd antibody rabbit pab/product/Sino Biological
    Average 95 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, 2021-06
    95/100 stars
      Buy from Supplier

    Image Search Results


    RU169 output clone diversity Using the SARS-CoV-2 RBD as the target of library panning and FACS selection for screen RU169 produced a high number of unique clones, indicating high, unexplored, diversity in the output.

    Journal: bioRxiv

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries

    doi: 10.1101/2020.07.27.224089

    Figure Lengend Snippet: RU169 output clone diversity Using the SARS-CoV-2 RBD as the target of library panning and FACS selection for screen RU169 produced a high number of unique clones, indicating high, unexplored, diversity in the output.

    Article Snippet: ACE2-S1 inhibition assayThe ability of RBD-binding antibodies to block the high-affinity interaction between SARS-CoV-2 RBD and human ACE2 protein was tested in a bead-binding assay.

    Techniques: FACS, Selection, Produced, Clone Assay

    BLI kinetics of selected scFv clones from the RU169 RBD screen. scFv were cloned into an AviTag™ biotinylation vector, as described in the Materials and Methods, expressed and purified by Ni-NTA resin. scFv were loaded onto a streptavidin BLI sensor and the association/dissociation kinetics of binding to soluble SARS-CoV-2 S1 trimer (100 nM) were measured using BLI. The K D of the scFvs for the S1 target ranged from 1 nM to 400 nM.

    Journal: bioRxiv

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries

    doi: 10.1101/2020.07.27.224089

    Figure Lengend Snippet: BLI kinetics of selected scFv clones from the RU169 RBD screen. scFv were cloned into an AviTag™ biotinylation vector, as described in the Materials and Methods, expressed and purified by Ni-NTA resin. scFv were loaded onto a streptavidin BLI sensor and the association/dissociation kinetics of binding to soluble SARS-CoV-2 S1 trimer (100 nM) were measured using BLI. The K D of the scFvs for the S1 target ranged from 1 nM to 400 nM.

    Article Snippet: ACE2-S1 inhibition assayThe ability of RBD-binding antibodies to block the high-affinity interaction between SARS-CoV-2 RBD and human ACE2 protein was tested in a bead-binding assay.

    Techniques: Clone Assay, Plasmid Preparation, Purification, Binding Assay

    Anti-RBD clones in IgG1 format form long-lived complexes with SARS-CoV-2 S1 trimer and potently inhibit the interaction with ACE2 in vitro . A. Dissociation kinetics of IgG1 anti-RBD clones from SARS-CoV-2 S1 trimer. Biotinylated SARS-CoV-2 S1 trimer was bound to a streptavidin BLI sensor. IgG1 anti-RBD clones were bound (100 nM) and the dissociation followed for 4 hours in PBS at 25°C. B. ACE2-S1 Dynabead assay with molar equivalents of mAb clones to S1 trimer.

    Journal: bioRxiv

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries

    doi: 10.1101/2020.07.27.224089

    Figure Lengend Snippet: Anti-RBD clones in IgG1 format form long-lived complexes with SARS-CoV-2 S1 trimer and potently inhibit the interaction with ACE2 in vitro . A. Dissociation kinetics of IgG1 anti-RBD clones from SARS-CoV-2 S1 trimer. Biotinylated SARS-CoV-2 S1 trimer was bound to a streptavidin BLI sensor. IgG1 anti-RBD clones were bound (100 nM) and the dissociation followed for 4 hours in PBS at 25°C. B. ACE2-S1 Dynabead assay with molar equivalents of mAb clones to S1 trimer.

    Article Snippet: ACE2-S1 inhibition assayThe ability of RBD-binding antibodies to block the high-affinity interaction between SARS-CoV-2 RBD and human ACE2 protein was tested in a bead-binding assay.

    Techniques: Clone Assay, In Vitro

    FACS strategy of screen RU167 for scFv inhibiting the SARS-CoV-2 RBD/ACE2 interaction The FACS-based screening strategy for screen RU167 to isolate antibodies that bound SARS-CoV-2 RBD and specifically inhibited co-binding of RBD to the human ACE2 protein. The viral RBD and the ACE2 protein were labeled with different fluorophores (A). Binding to cells expressing scFv clones that bound RBD and blocking the ACE2-binding site (B) would be observed and gated positively for in the FACS plot for events which were RBD-dye HIGH and ACE2-dye LOW (C).

    Journal: bioRxiv

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries

    doi: 10.1101/2020.07.27.224089

    Figure Lengend Snippet: FACS strategy of screen RU167 for scFv inhibiting the SARS-CoV-2 RBD/ACE2 interaction The FACS-based screening strategy for screen RU167 to isolate antibodies that bound SARS-CoV-2 RBD and specifically inhibited co-binding of RBD to the human ACE2 protein. The viral RBD and the ACE2 protein were labeled with different fluorophores (A). Binding to cells expressing scFv clones that bound RBD and blocking the ACE2-binding site (B) would be observed and gated positively for in the FACS plot for events which were RBD-dye HIGH and ACE2-dye LOW (C).

    Article Snippet: ACE2-S1 inhibition assayThe ability of RBD-binding antibodies to block the high-affinity interaction between SARS-CoV-2 RBD and human ACE2 protein was tested in a bead-binding assay.

    Techniques: FACS, Binding Assay, Labeling, Expressing, Clone Assay, Blocking Assay

    BLI kinetics of anti-RBD diabodies AviTag™ biotinylated SARS-CoV-2 S1 trimer was loaded onto a BLI sensor and the association/dissociation kinetics of binding to anti-RBD diabodies (100 nM) were measured using BLI. The K D s of the dbs to the S1 target ranged from 84 pM to 1 nM.

    Journal: bioRxiv

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries

    doi: 10.1101/2020.07.27.224089

    Figure Lengend Snippet: BLI kinetics of anti-RBD diabodies AviTag™ biotinylated SARS-CoV-2 S1 trimer was loaded onto a BLI sensor and the association/dissociation kinetics of binding to anti-RBD diabodies (100 nM) were measured using BLI. The K D s of the dbs to the S1 target ranged from 84 pM to 1 nM.

    Article Snippet: ACE2-S1 inhibition assayThe ability of RBD-binding antibodies to block the high-affinity interaction between SARS-CoV-2 RBD and human ACE2 protein was tested in a bead-binding assay.

    Techniques: Binding Assay

    Cytometry plots of ACE2-S1 Dynabead assay of anti-RBD diabodies The degree of inhibition of the ACE2 and SARS-CoV-2 S1 trimer interaction by stoichiometric amounts of anti-RBD diabodies was determined using a Dynabead assay as described in the Materials and Methods. The degree of bead fluorescence was indicative of the amount of dye-labeled S1 trimer that was bound to ACE2. Inhibition of the interaction by anti-RBD diabodies resulted in a reduction in fluorescence. The first panel is the SSC/FSC indicating the P1 gating of beads. The second panel is the biotin-blocked control (no ACE2/S1 interaction) and the third panel is the no anti-RBD control (maximum ACE2/S1 interaction. Each subsequent row represents a db clone at 1:1, 5:1 and 10:1 stoichiometric ratios to the soluble SARS-CoV-2 S1 trimer. The data are summarized graphically in Figure 3 .

    Journal: bioRxiv

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries

    doi: 10.1101/2020.07.27.224089

    Figure Lengend Snippet: Cytometry plots of ACE2-S1 Dynabead assay of anti-RBD diabodies The degree of inhibition of the ACE2 and SARS-CoV-2 S1 trimer interaction by stoichiometric amounts of anti-RBD diabodies was determined using a Dynabead assay as described in the Materials and Methods. The degree of bead fluorescence was indicative of the amount of dye-labeled S1 trimer that was bound to ACE2. Inhibition of the interaction by anti-RBD diabodies resulted in a reduction in fluorescence. The first panel is the SSC/FSC indicating the P1 gating of beads. The second panel is the biotin-blocked control (no ACE2/S1 interaction) and the third panel is the no anti-RBD control (maximum ACE2/S1 interaction. Each subsequent row represents a db clone at 1:1, 5:1 and 10:1 stoichiometric ratios to the soluble SARS-CoV-2 S1 trimer. The data are summarized graphically in Figure 3 .

    Article Snippet: ACE2-S1 inhibition assayThe ability of RBD-binding antibodies to block the high-affinity interaction between SARS-CoV-2 RBD and human ACE2 protein was tested in a bead-binding assay.

    Techniques: Cytometry, Inhibition, Fluorescence, Labeling

    Results from the Adarza Ziva system for pre-COVID-19 serum samples and single-donor samples from convalescent COVID-19 (PCR-positive) subjects. Pre-COVID-19 single-donor results were averaged (blue bars). Black bars indicate threshold positive values, calculated as two standard deviations above the average negative (pre-COVID-19) signal. Red bars indicate PCR+ individuals yielding signals below the threshold on all SARS-CoV-2 antigens, while green bars indicate signals from single-donor convalescent COVID-19 samples with at least one SARS-CoV-2 antigen response above threshold.

    Journal: bioRxiv

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

    doi: 10.1101/2020.06.15.153064

    Figure Lengend Snippet: Results from the Adarza Ziva system for pre-COVID-19 serum samples and single-donor samples from convalescent COVID-19 (PCR-positive) subjects. Pre-COVID-19 single-donor results were averaged (blue bars). Black bars indicate threshold positive values, calculated as two standard deviations above the average negative (pre-COVID-19) signal. Red bars indicate PCR+ individuals yielding signals below the threshold on all SARS-CoV-2 antigens, while green bars indicate signals from single-donor convalescent COVID-19 samples with at least one SARS-CoV-2 antigen response above threshold.

    Article Snippet: Calculated limits of detection for these data were 43.3 ng/mL (SARS-CoV-2 S1 + S2 ECD), 40.7 ng/mL (SARS-CoV-2 S1), and 25.1 ng/mL (SARS-CoV-2 RBD).

    Techniques: Polymerase Chain Reaction

    AIR assay for antibodies to respiratory viruses. For each antigen, six replicate spots are printed in two different locations on the chip. Each group of six spots is surrounded by negative control reference spots (anti-FITC). Blank (background) areas are included as additional negative controls. Key: 1: human coronavirus (HKU isolate) spike glycoprotein, aa 1-760; 2: MERS-CoV spike glycoprotein, S1 domain; 3: MERS-CoV spike glycoprotein, receptor binding domain (RBD); 4: SARS-CoV spike glycoprotein, S1 domain; 5: SARS-CoV spike glycoprotein, RBD; 6: SARS-CoV-2 spike glycoprotein, S1+S2 ECD; 7: SARS-CoV-2 spike glycoprotein, S2 ECD; 8: SARS-CoV-2 spike glycoprotein, S1 domain; 9: SARS-CoV-2 spike glycoprotein, RBD; 10: human coronavirus (HCoV-229E isolate) spike glycoprotein, S1+S2 ECD; 11: human coronavirus (HCoV-OC43 isolate) spike glycoprotein, S1+S2 ECD; 12: influenza B/Brisbane/2008 hemagglutinin; 13: influenza A/California/2009 (H1N1) hemagglutinin; 14: influenza A/Wisconsin/2005 (H3N2) influenza. F1 , F2 , and F3 are derived from spotting three different dilutions of anti-FITC. The image at right is a representative array exposed to Pooled Normal Human Serum (PNHS) at a 1:4 dilution.

    Journal: bioRxiv

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

    doi: 10.1101/2020.06.15.153064

    Figure Lengend Snippet: AIR assay for antibodies to respiratory viruses. For each antigen, six replicate spots are printed in two different locations on the chip. Each group of six spots is surrounded by negative control reference spots (anti-FITC). Blank (background) areas are included as additional negative controls. Key: 1: human coronavirus (HKU isolate) spike glycoprotein, aa 1-760; 2: MERS-CoV spike glycoprotein, S1 domain; 3: MERS-CoV spike glycoprotein, receptor binding domain (RBD); 4: SARS-CoV spike glycoprotein, S1 domain; 5: SARS-CoV spike glycoprotein, RBD; 6: SARS-CoV-2 spike glycoprotein, S1+S2 ECD; 7: SARS-CoV-2 spike glycoprotein, S2 ECD; 8: SARS-CoV-2 spike glycoprotein, S1 domain; 9: SARS-CoV-2 spike glycoprotein, RBD; 10: human coronavirus (HCoV-229E isolate) spike glycoprotein, S1+S2 ECD; 11: human coronavirus (HCoV-OC43 isolate) spike glycoprotein, S1+S2 ECD; 12: influenza B/Brisbane/2008 hemagglutinin; 13: influenza A/California/2009 (H1N1) hemagglutinin; 14: influenza A/Wisconsin/2005 (H3N2) influenza. F1 , F2 , and F3 are derived from spotting three different dilutions of anti-FITC. The image at right is a representative array exposed to Pooled Normal Human Serum (PNHS) at a 1:4 dilution.

    Article Snippet: Calculated limits of detection for these data were 43.3 ng/mL (SARS-CoV-2 S1 + S2 ECD), 40.7 ng/mL (SARS-CoV-2 S1), and 25.1 ng/mL (SARS-CoV-2 RBD).

    Techniques: Chromatin Immunoprecipitation, Negative Control, Binding Assay, Derivative Assay

    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.

    Journal: bioRxiv

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

    doi: 10.1101/2020.06.15.153064

    Figure Lengend 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.

    Article Snippet: Calculated limits of detection for these data were 43.3 ng/mL (SARS-CoV-2 S1 + S2 ECD), 40.7 ng/mL (SARS-CoV-2 S1), and 25.1 ng/mL (SARS-CoV-2 RBD).

    Techniques: Enzyme-linked Immunosorbent Assay

    Representative AIR array images (100 ms exposures) of (A) 5% FBS; (B) 10% PNHS; (C) a negative single-donor sample, and (D) one convalescent serum sample. Strong responses to SARS-CoV-2 antigens are readily observed in (D), but not in (A), (B), or (C). In each case, samples were diluted 1:20 in Adarza diluent, and incubated with the arrays overnight at 4 °C. See Figure 1 for key to the array. All arrays in this figure were imaged at an exposure of 100 ms.

    Journal: bioRxiv

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

    doi: 10.1101/2020.06.15.153064

    Figure Lengend Snippet: Representative AIR array images (100 ms exposures) of (A) 5% FBS; (B) 10% PNHS; (C) a negative single-donor sample, and (D) one convalescent serum sample. Strong responses to SARS-CoV-2 antigens are readily observed in (D), but not in (A), (B), or (C). In each case, samples were diluted 1:20 in Adarza diluent, and incubated with the arrays overnight at 4 °C. See Figure 1 for key to the array. All arrays in this figure were imaged at an exposure of 100 ms.

    Article Snippet: Calculated limits of detection for these data were 43.3 ng/mL (SARS-CoV-2 S1 + S2 ECD), 40.7 ng/mL (SARS-CoV-2 S1), and 25.1 ng/mL (SARS-CoV-2 RBD).

    Techniques: Incubation

    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.

    Journal: bioRxiv

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

    doi: 10.1101/2020.06.15.153064

    Figure Lengend 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.

    Article Snippet: Calculated limits of detection for these data were 43.3 ng/mL (SARS-CoV-2 S1 + S2 ECD), 40.7 ng/mL (SARS-CoV-2 S1), and 25.1 ng/mL (SARS-CoV-2 RBD).

    Techniques: Titration, Concentration Assay, Standard Deviation

    ELISA ( x -axis) vs. LFRET ( y -axis) results by disease severity. ( a ) Anti-NP IgA ELISA vs. anti-NP LFRET (N = 81, R = 0.25). ( b ) anti-NP IgG ELISA vs. anti-NP LFRET (N = 129, R = 0.62). ( c ) anti-NP IgM ELISA vs. anti-NP LFRET (N = 81, R = 0.13). ( d ) anti-SP IgA ELISA vs. anti-SP LFRET (N = 129, R = 0.53). ( e ) anti-SP IgG ELISA vs. anti-SP LFRET (N = 129, R = 0.62). ( f ) anti-SP IgM ELISA vs. anti-SP LFRET (N = 81, R = 0.56). Color of the dot indicates SARS-CoV-2 PCR result and disease severity: cyan = PCR negative; yellow = non-hospitalized, PCR-positive; red = non-ICU hospitalized, PCR positive; black = hospitalized in ICU, PCR positive. Horizontal and vertical black lines indicate LFRET and ELISA cutoffs. On the x -axis, ELISA absorbance on a logarithmic scale and on the y -axis, LFRET signal on a logarithmic scale. SP = spike glycoprotein. NP = nucleoprotein. LFRET = protein L–based time-resolved Förster resonance energy transfer immunoassay. ELISA = enzyme immunoassay. R = Pearson’s correlation coefficient.

    Journal: Viruses

    Article Title: A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2

    doi: 10.3390/v13020143

    Figure Lengend Snippet: ELISA ( x -axis) vs. LFRET ( y -axis) results by disease severity. ( a ) Anti-NP IgA ELISA vs. anti-NP LFRET (N = 81, R = 0.25). ( b ) anti-NP IgG ELISA vs. anti-NP LFRET (N = 129, R = 0.62). ( c ) anti-NP IgM ELISA vs. anti-NP LFRET (N = 81, R = 0.13). ( d ) anti-SP IgA ELISA vs. anti-SP LFRET (N = 129, R = 0.53). ( e ) anti-SP IgG ELISA vs. anti-SP LFRET (N = 129, R = 0.62). ( f ) anti-SP IgM ELISA vs. anti-SP LFRET (N = 81, R = 0.56). Color of the dot indicates SARS-CoV-2 PCR result and disease severity: cyan = PCR negative; yellow = non-hospitalized, PCR-positive; red = non-ICU hospitalized, PCR positive; black = hospitalized in ICU, PCR positive. Horizontal and vertical black lines indicate LFRET and ELISA cutoffs. On the x -axis, ELISA absorbance on a logarithmic scale and on the y -axis, LFRET signal on a logarithmic scale. SP = spike glycoprotein. NP = nucleoprotein. LFRET = protein L–based time-resolved Förster resonance energy transfer immunoassay. ELISA = enzyme immunoassay. R = Pearson’s correlation coefficient.

    Article Snippet: At 48 h, the medium was analyzed for the presence of SARS-CoV-2 SP by dot blotting; briefly via drying 2.5 µL of the supernatant onto a nitrocellulose membrane, which then was blocked (3% skim milk in Tris-buffered saline with 0.05% Tween-20), washed, probed with rabbit anti-RBD (40592-T62, Sino Biological, Beijing, China), washed, probed with anti-rabbit IRDye800 (LI-COR Biosciences, Lincoln, NE, USA), washed, and read using Odyssey Infrared Imaging System (LI-COR Biosciences).

    Techniques: Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Förster Resonance Energy Transfer

    Microneutralization vs. LFRET and ELISA. Microneutralization titers are on the x -axis and LFRET signal or ELISA absorbance on the y -axis. Logarithmic scale is used on both axes. ( a ) Microneutralization titer vs. anti-SP LFRET signal (N = 107, ρ = 0.87). ( b – d ) Microneutralization titer vs. anti-SP IgG, IgA and IgM ELISA (N = 107, 107 and 67, ρ = 0.68, 0.86 and 0.81). ( e ) Microneutralization titer vs. anti-NP LFRET signal (N = 107, ρ = 0.83). ( f – h ) Microneutralization titer vs. anti-NP IgG, IgA and IgM ELISA (N = 107, 67 and 67, ρ = 0.81, 0.69 and 0.61). Color of the dots indicate SARS-CoV-2 PCR result and disease severity: cyan = PCR negative; yellow = non-hospitalized, PCR-positive; red = non-ICU hospitalized, PCR positive; black = hospitalized in ICU, PCR positive. Horizontal black lines indicate LFRET/ELISA cutoffs. SP = spike glycoprotein. NP = nucleoprotein. LFRET = protein L–based time-resolved Förster resonance energy transfer immunoassay. ELISA = enzyme immunoassay. ρ = Spearman’s rank correlation coefficient.

    Journal: Viruses

    Article Title: A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2

    doi: 10.3390/v13020143

    Figure Lengend Snippet: Microneutralization vs. LFRET and ELISA. Microneutralization titers are on the x -axis and LFRET signal or ELISA absorbance on the y -axis. Logarithmic scale is used on both axes. ( a ) Microneutralization titer vs. anti-SP LFRET signal (N = 107, ρ = 0.87). ( b – d ) Microneutralization titer vs. anti-SP IgG, IgA and IgM ELISA (N = 107, 107 and 67, ρ = 0.68, 0.86 and 0.81). ( e ) Microneutralization titer vs. anti-NP LFRET signal (N = 107, ρ = 0.83). ( f – h ) Microneutralization titer vs. anti-NP IgG, IgA and IgM ELISA (N = 107, 67 and 67, ρ = 0.81, 0.69 and 0.61). Color of the dots indicate SARS-CoV-2 PCR result and disease severity: cyan = PCR negative; yellow = non-hospitalized, PCR-positive; red = non-ICU hospitalized, PCR positive; black = hospitalized in ICU, PCR positive. Horizontal black lines indicate LFRET/ELISA cutoffs. SP = spike glycoprotein. NP = nucleoprotein. LFRET = protein L–based time-resolved Förster resonance energy transfer immunoassay. ELISA = enzyme immunoassay. ρ = Spearman’s rank correlation coefficient.

    Article Snippet: At 48 h, the medium was analyzed for the presence of SARS-CoV-2 SP by dot blotting; briefly via drying 2.5 µL of the supernatant onto a nitrocellulose membrane, which then was blocked (3% skim milk in Tris-buffered saline with 0.05% Tween-20), washed, probed with rabbit anti-RBD (40592-T62, Sino Biological, Beijing, China), washed, probed with anti-rabbit IRDye800 (LI-COR Biosciences, Lincoln, NE, USA), washed, and read using Odyssey Infrared Imaging System (LI-COR Biosciences).

    Techniques: Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction, Förster Resonance Energy Transfer

    Simplified protocol for SARS-CoV-2 NP and SP LFRET assay. Eu-NP/-SP = Europium-labeled nucleoprotein/spike glycoprotein. AF-L = Alexa Fluor™ 647 -labeled protein L. TR-FRET = time-resolved Förster resonance energy transfer. RT = room temperature. TBS+BSA (50 mM Tris-HCl, 150 mM NaCl, pH 7.4, 0.2% BSA) was used for all dilutions. On-plate dilutions were 5 nM Eu-NP/500 nM AF-L/serum 1/25 for anti-NP and 5 nM Eu-SP/250 nM AF-L/serum 1/100 for anti-SP LFRET. For further details see the prior publication [ 5 ].

    Journal: Viruses

    Article Title: A 10-Minute “Mix and Read” Antibody Assay for SARS-CoV-2

    doi: 10.3390/v13020143

    Figure Lengend Snippet: Simplified protocol for SARS-CoV-2 NP and SP LFRET assay. Eu-NP/-SP = Europium-labeled nucleoprotein/spike glycoprotein. AF-L = Alexa Fluor™ 647 -labeled protein L. TR-FRET = time-resolved Förster resonance energy transfer. RT = room temperature. TBS+BSA (50 mM Tris-HCl, 150 mM NaCl, pH 7.4, 0.2% BSA) was used for all dilutions. On-plate dilutions were 5 nM Eu-NP/500 nM AF-L/serum 1/25 for anti-NP and 5 nM Eu-SP/250 nM AF-L/serum 1/100 for anti-SP LFRET. For further details see the prior publication [ 5 ].

    Article Snippet: At 48 h, the medium was analyzed for the presence of SARS-CoV-2 SP by dot blotting; briefly via drying 2.5 µL of the supernatant onto a nitrocellulose membrane, which then was blocked (3% skim milk in Tris-buffered saline with 0.05% Tween-20), washed, probed with rabbit anti-RBD (40592-T62, Sino Biological, Beijing, China), washed, probed with anti-rabbit IRDye800 (LI-COR Biosciences, Lincoln, NE, USA), washed, and read using Odyssey Infrared Imaging System (LI-COR Biosciences).

    Techniques: Labeling, Förster Resonance Energy Transfer

    Inducible Bronchus Associated Lymphoid Tissues (iBALT) formation upon MVA/S and MVA/S1 vaccination. Frozen lung sections from vaccinated mice were either stained for H E to analyze tissue structure and formation of iBALT aggregates (A), or immunofluorescence stained to visualize B cell and T cell (B) forming B cell follicle like structure (iBALT) induced by MVA/S vaccination given via i.m. route (right panel), and compared with unvaccinated control mice (left panel). Total number of iBALT like structures visualized in each section per mice was quantified and compared between the groups (C). The p value was calculated using non parametric mann-whitney test. (D) Lung immune responses in bronchoalveolar lavage (BAL) samples collected after euthanizations (three weeks post-boost) were measured using ELISA. SARS-CoV-2 S protein-specific binding IgG and IgA antibodies measured, and titters were presented in column graphs. The data represent mean responses in each group (n = 5) ± SEM.

    Journal: bioRxiv

    Article Title: Modified Vaccinia Ankara Based SARS-CoV-2 Vaccine Expressing Full-Length Spike Induces Strong Neutralizing Antibody Response

    doi: 10.1101/2020.06.27.175166

    Figure Lengend Snippet: Inducible Bronchus Associated Lymphoid Tissues (iBALT) formation upon MVA/S and MVA/S1 vaccination. Frozen lung sections from vaccinated mice were either stained for H E to analyze tissue structure and formation of iBALT aggregates (A), or immunofluorescence stained to visualize B cell and T cell (B) forming B cell follicle like structure (iBALT) induced by MVA/S vaccination given via i.m. route (right panel), and compared with unvaccinated control mice (left panel). Total number of iBALT like structures visualized in each section per mice was quantified and compared between the groups (C). The p value was calculated using non parametric mann-whitney test. (D) Lung immune responses in bronchoalveolar lavage (BAL) samples collected after euthanizations (three weeks post-boost) were measured using ELISA. SARS-CoV-2 S protein-specific binding IgG and IgA antibodies measured, and titters were presented in column graphs. The data represent mean responses in each group (n = 5) ± SEM.

    Article Snippet: Proteins were transferred to a nitrocellulose membrane, blocked with 1% casein blocker overnight (Cat#1610782 Biorad), and incubated for 1 h at room temperature with anti-SARS-CoV-2 spike mouse mAb (Cat # GTX632604, GeneTex) for MVA/S and rabbit SARS-CoV-2 RBD polyclonal antibody (Cat# 40592-T62, Sino Biological) for MVA/S1 diluted 1:2500 in blocking buffer, respectively.

    Techniques: Mouse Assay, Staining, Immunofluorescence, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay, Binding Assay

    Neutralizing activity against SARS-CoV-2. (A) Percent neutralization of SARS-CoV-2 virus expressing GFP. Serum collected from the naïve animals used as negative controls. (B) Neutralization titer against SARS-CoV-2 virus expressing GFP. (C, D) Correlations between neutralization titer and ELISA binding titer.

    Journal: bioRxiv

    Article Title: Modified Vaccinia Ankara Based SARS-CoV-2 Vaccine Expressing Full-Length Spike Induces Strong Neutralizing Antibody Response

    doi: 10.1101/2020.06.27.175166

    Figure Lengend Snippet: Neutralizing activity against SARS-CoV-2. (A) Percent neutralization of SARS-CoV-2 virus expressing GFP. Serum collected from the naïve animals used as negative controls. (B) Neutralization titer against SARS-CoV-2 virus expressing GFP. (C, D) Correlations between neutralization titer and ELISA binding titer.

    Article Snippet: Proteins were transferred to a nitrocellulose membrane, blocked with 1% casein blocker overnight (Cat#1610782 Biorad), and incubated for 1 h at room temperature with anti-SARS-CoV-2 spike mouse mAb (Cat # GTX632604, GeneTex) for MVA/S and rabbit SARS-CoV-2 RBD polyclonal antibody (Cat# 40592-T62, Sino Biological) for MVA/S1 diluted 1:2500 in blocking buffer, respectively.

    Techniques: Activity Assay, Neutralization, Expressing, Enzyme-linked Immunosorbent Assay, Binding Assay

    Analyzing SARS-CoV-2 RBD and S1 proteins affinities to human ACE2 (hACE2) proteins using biolayer interferometry (BLI). (A) Bio-Layer Interferometry sensograms of the binding of SARS-CoV-2 S1 and RBD proteins to immobilized Fc-human ACE2, after incubation of the analytes at 25°C for 0and 60 minutes. The traces represent BLI response curves for SARS-CoV-2 proteins serially diluted from 800nM to 12.5nM, as indicated. Dotted lines show raw response values, while bold solid lines show the fitted trace. Association and dissociation phases were monitored for 300s and 600s, respectively. The data was globally fit using a 1:1 binding model to estimate binding affinity. (B) Binding affinity specifications of S1 and RBD proteins against hu-ACE2.

    Journal: bioRxiv

    Article Title: Modified Vaccinia Ankara Based SARS-CoV-2 Vaccine Expressing Full-Length Spike Induces Strong Neutralizing Antibody Response

    doi: 10.1101/2020.06.27.175166

    Figure Lengend Snippet: Analyzing SARS-CoV-2 RBD and S1 proteins affinities to human ACE2 (hACE2) proteins using biolayer interferometry (BLI). (A) Bio-Layer Interferometry sensograms of the binding of SARS-CoV-2 S1 and RBD proteins to immobilized Fc-human ACE2, after incubation of the analytes at 25°C for 0and 60 minutes. The traces represent BLI response curves for SARS-CoV-2 proteins serially diluted from 800nM to 12.5nM, as indicated. Dotted lines show raw response values, while bold solid lines show the fitted trace. Association and dissociation phases were monitored for 300s and 600s, respectively. The data was globally fit using a 1:1 binding model to estimate binding affinity. (B) Binding affinity specifications of S1 and RBD proteins against hu-ACE2.

    Article Snippet: Proteins were transferred to a nitrocellulose membrane, blocked with 1% casein blocker overnight (Cat#1610782 Biorad), and incubated for 1 h at room temperature with anti-SARS-CoV-2 spike mouse mAb (Cat # GTX632604, GeneTex) for MVA/S and rabbit SARS-CoV-2 RBD polyclonal antibody (Cat# 40592-T62, Sino Biological) for MVA/S1 diluted 1:2500 in blocking buffer, respectively.

    Techniques: Binding Assay, Incubation

    Antibody responses induced by MVA/S or MVA/S1 in mice. BALB/c mice were immunized on week 0 and 3 with recombinant MVAs expressing either S (MVA/S) (n=5) or S1 (MVA/S1) (n=5) in a prime-boost strategy. Unvaccinated (naïve) animals served as controls (n=5). (A) Binding IgG antibody response for individual proteins measured using ELISA at two weeks after boost. (B) Endpoint IgG titers against SARS-CoV-2 RBD, S1 and S measured at week 2 after immunization. The data show mean response in each group (n = 5) ± SEM. (C) Binding antibody response determined using Luminex assay at 3 weeks post boost. The pie graphs show the relative proportions of binding to three proteins in each group. (D) IgG subclass and soluble Fc receptor binding analysis of RBD and S1 specific IgG measured using the Luminex assay. Raw values are presented as in mean fluorescence intensity (MFI) in bar graph. The data represent mean responses in each group (n = 5) ± SEM.

    Journal: bioRxiv

    Article Title: Modified Vaccinia Ankara Based SARS-CoV-2 Vaccine Expressing Full-Length Spike Induces Strong Neutralizing Antibody Response

    doi: 10.1101/2020.06.27.175166

    Figure Lengend Snippet: Antibody responses induced by MVA/S or MVA/S1 in mice. BALB/c mice were immunized on week 0 and 3 with recombinant MVAs expressing either S (MVA/S) (n=5) or S1 (MVA/S1) (n=5) in a prime-boost strategy. Unvaccinated (naïve) animals served as controls (n=5). (A) Binding IgG antibody response for individual proteins measured using ELISA at two weeks after boost. (B) Endpoint IgG titers against SARS-CoV-2 RBD, S1 and S measured at week 2 after immunization. The data show mean response in each group (n = 5) ± SEM. (C) Binding antibody response determined using Luminex assay at 3 weeks post boost. The pie graphs show the relative proportions of binding to three proteins in each group. (D) IgG subclass and soluble Fc receptor binding analysis of RBD and S1 specific IgG measured using the Luminex assay. Raw values are presented as in mean fluorescence intensity (MFI) in bar graph. The data represent mean responses in each group (n = 5) ± SEM.

    Article Snippet: Proteins were transferred to a nitrocellulose membrane, blocked with 1% casein blocker overnight (Cat#1610782 Biorad), and incubated for 1 h at room temperature with anti-SARS-CoV-2 spike mouse mAb (Cat # GTX632604, GeneTex) for MVA/S and rabbit SARS-CoV-2 RBD polyclonal antibody (Cat# 40592-T62, Sino Biological) for MVA/S1 diluted 1:2500 in blocking buffer, respectively.

    Techniques: Mouse Assay, Recombinant, Expressing, Binding Assay, Enzyme-linked Immunosorbent Assay, Luminex, Fluorescence