hku1  (ATCC)


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    ATCC hku1
    Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and <t>-HKU1</t> (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.
    Hku1, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Characterization of spike processing and entry mechanisms of seasonal human coronaviruses NL63, 229E and HKU1"

    Article Title: Characterization of spike processing and entry mechanisms of seasonal human coronaviruses NL63, 229E and HKU1

    Journal: bioRxiv

    doi: 10.1101/2024.04.12.589332

    Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and -HKU1 (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.
    Figure Legend Snippet: Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and -HKU1 (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.

    Techniques Used: Infection, Comparison, Mutagenesis, Software, Western Blot

    (A) Chloroquine sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (B) Camostat mesylate sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (C) Chloroquine sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (D) Ammonium chloride sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (E) Chloroquine sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2 cells. (F) Camostat mesylate sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2-TMPRSS2 cells respectively. For all experiments, cells were pretreated with indicated inhibitor for 2 h prior to pseudovirus infection in the presence of inhibitor for a period of 48 h. X-axis indicates log concentration of inhibitor. Y axis indicates percentage inhibition compared to pseudovirus infection without inhibitor treatment. Results shown are representative of three independent experiments.
    Figure Legend Snippet: (A) Chloroquine sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (B) Camostat mesylate sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (C) Chloroquine sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (D) Ammonium chloride sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (E) Chloroquine sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2 cells. (F) Camostat mesylate sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2-TMPRSS2 cells respectively. For all experiments, cells were pretreated with indicated inhibitor for 2 h prior to pseudovirus infection in the presence of inhibitor for a period of 48 h. X-axis indicates log concentration of inhibitor. Y axis indicates percentage inhibition compared to pseudovirus infection without inhibitor treatment. Results shown are representative of three independent experiments.

    Techniques Used: Infection, Concentration Assay, Inhibition

    (A) Cellular TMPRSS2 is a type II transmembrane protease comprising N-terminal transmembrane (TM) domain, Low Density Lipoprotein Receptor domain A (LDLRA), Class A Scavenger Receptor Cysteine-Rich (SRCR) domain and a C-terminal trypsin-like serine protease (SP) domain with a canonical H296-D345-S441 catalytic triad. TMPRSS2 with an N-terminal Flag tag, LDLRA domain of TMPRSS2 with an N-terminal Flag tag, LDLRA and SRCR domains of TMPRSS2 with an N-terminal Flag tag, and TMPRSS2 proteins with alanine substitutions in the indicated active site residues of catalytic triad are shown. (B) Western Blotting of Flag-tagged TMPRSS2 proteins in 293T cells transfected with indicated plasmids. Lane 1 is Marker, Lane 2 showed TMPRSS2 expressed as full-length form (∼54KD) and cleavage products (∼20-25KD, 37-40KD) due to autoactivation. Lane 3-5 showed TMPRSS2 proteins lacking protease catalytic acitivity expressed mainly as full-length forms (∼54KD). Lanes 6 and show LDLRA and LDLRA-SRCR domains expressed as 25KD and 37KD forms. Lane shows Flag-tagged ACE2 (∼110KD) as a control. (C) Infectivity of VSV pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of VSV pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue, and green bars respectively (D) Infectivity of SARSCoV-2 pseudovirses in 293T-ACE2 cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of SARS-CoV-2 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively (E) Infectivity of HCoV-HKU1 pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of HCoV-HKU1 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively. The tests for two-group comparison (infectivity without inhibitor vs infectivity with inhibitor) were analyzed using GraphPad Prism software. Asterisk* denotes significance: *: p ≤ 0.05; **: p ≤ 0.01; ****: p ≤ 0.0001 .
    Figure Legend Snippet: (A) Cellular TMPRSS2 is a type II transmembrane protease comprising N-terminal transmembrane (TM) domain, Low Density Lipoprotein Receptor domain A (LDLRA), Class A Scavenger Receptor Cysteine-Rich (SRCR) domain and a C-terminal trypsin-like serine protease (SP) domain with a canonical H296-D345-S441 catalytic triad. TMPRSS2 with an N-terminal Flag tag, LDLRA domain of TMPRSS2 with an N-terminal Flag tag, LDLRA and SRCR domains of TMPRSS2 with an N-terminal Flag tag, and TMPRSS2 proteins with alanine substitutions in the indicated active site residues of catalytic triad are shown. (B) Western Blotting of Flag-tagged TMPRSS2 proteins in 293T cells transfected with indicated plasmids. Lane 1 is Marker, Lane 2 showed TMPRSS2 expressed as full-length form (∼54KD) and cleavage products (∼20-25KD, 37-40KD) due to autoactivation. Lane 3-5 showed TMPRSS2 proteins lacking protease catalytic acitivity expressed mainly as full-length forms (∼54KD). Lanes 6 and show LDLRA and LDLRA-SRCR domains expressed as 25KD and 37KD forms. Lane shows Flag-tagged ACE2 (∼110KD) as a control. (C) Infectivity of VSV pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of VSV pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue, and green bars respectively (D) Infectivity of SARSCoV-2 pseudovirses in 293T-ACE2 cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of SARS-CoV-2 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively (E) Infectivity of HCoV-HKU1 pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of HCoV-HKU1 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively. The tests for two-group comparison (infectivity without inhibitor vs infectivity with inhibitor) were analyzed using GraphPad Prism software. Asterisk* denotes significance: *: p ≤ 0.05; **: p ≤ 0.01; ****: p ≤ 0.0001 .

    Techniques Used: FLAG-tag, Western Blot, Transfection, Marker, Infection, Comparison, Software

    (A) Western blotting of HCoV-HKU1 spike in Anti-Flag Ab pull downs immunoprecipitated by indicated TMPRSS2 proteins with a Flag tag at the N-terminus. HCoV-HKU1 spike is probed using polyclonal serum. (B) Sample input employed for immunoprecipitation. TMPRSS2 proteins Flag tagged in the N-terminus are probed using anti-Flag antibody. Β-actin is loading control. (C) HCoV-HKU1 spike is expressed as both full length S0 and furin processed S1 forms. HCoV-HKU1-ΔFCS, which expresses only the full length S0 form is shown for size comparison.
    Figure Legend Snippet: (A) Western blotting of HCoV-HKU1 spike in Anti-Flag Ab pull downs immunoprecipitated by indicated TMPRSS2 proteins with a Flag tag at the N-terminus. HCoV-HKU1 spike is probed using polyclonal serum. (B) Sample input employed for immunoprecipitation. TMPRSS2 proteins Flag tagged in the N-terminus are probed using anti-Flag antibody. Β-actin is loading control. (C) HCoV-HKU1 spike is expressed as both full length S0 and furin processed S1 forms. HCoV-HKU1-ΔFCS, which expresses only the full length S0 form is shown for size comparison.

    Techniques Used: Western Blot, Immunoprecipitation, FLAG-tag, Comparison

    hcov hku1  (ATCC)


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    ATCC hcov hku1
    Hcov Hku1, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hcov hku1 c n5  (ATCC)


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    ATCC hcov hku1 c n5
    Hcov Hku1 C N5, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hku1  (ATCC)


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    ATCC hku1
    Hku1, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hku1  (ATCC)


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    ATCC hku1
    Evaluation of RHAM specificity. ( A ) The specificity of RHAM targeting Orf1ab was evaluated by testing the nucleic acids of other respiratory pathogens, ( B ) The specificity of RHAM targeting N gene was evaluated by testing the nucleic acids of other respiratory pathogens. All pathogens including coronavirus <t>(HKU1,</t> OC43, NL63, 229E), SARS coronavirus, MERS coronavirus, influenza A H1N1, H5N1, H7N9, and H9N2, influenza B virus, respiratory syncytial virus types A and B, human parainfluenza virus type II, adenovirus types 3 and 7, enterovirus EV71, Mycoplasma pneumoniae , Epstein-Barr virus, human cytomegalovirus, and Mycobacterium tuberculosis . 10 μL of nucleic acid was added to the RHAM reaction mixture (25 μL reaction volume).
    Hku1, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Rapid and accurate detection of SARS-CoV-2 using the RHAM technology"

    Article Title: Rapid and accurate detection of SARS-CoV-2 using the RHAM technology

    Journal: Scientific Reports

    doi: 10.1038/s41598-023-49733-7

    Evaluation of RHAM specificity. ( A ) The specificity of RHAM targeting Orf1ab was evaluated by testing the nucleic acids of other respiratory pathogens, ( B ) The specificity of RHAM targeting N gene was evaluated by testing the nucleic acids of other respiratory pathogens. All pathogens including coronavirus (HKU1, OC43, NL63, 229E), SARS coronavirus, MERS coronavirus, influenza A H1N1, H5N1, H7N9, and H9N2, influenza B virus, respiratory syncytial virus types A and B, human parainfluenza virus type II, adenovirus types 3 and 7, enterovirus EV71, Mycoplasma pneumoniae , Epstein-Barr virus, human cytomegalovirus, and Mycobacterium tuberculosis . 10 μL of nucleic acid was added to the RHAM reaction mixture (25 μL reaction volume).
    Figure Legend Snippet: Evaluation of RHAM specificity. ( A ) The specificity of RHAM targeting Orf1ab was evaluated by testing the nucleic acids of other respiratory pathogens, ( B ) The specificity of RHAM targeting N gene was evaluated by testing the nucleic acids of other respiratory pathogens. All pathogens including coronavirus (HKU1, OC43, NL63, 229E), SARS coronavirus, MERS coronavirus, influenza A H1N1, H5N1, H7N9, and H9N2, influenza B virus, respiratory syncytial virus types A and B, human parainfluenza virus type II, adenovirus types 3 and 7, enterovirus EV71, Mycoplasma pneumoniae , Epstein-Barr virus, human cytomegalovirus, and Mycobacterium tuberculosis . 10 μL of nucleic acid was added to the RHAM reaction mixture (25 μL reaction volume).

    Techniques Used: Virus

    hcov hku1 c n5  (ATCC)


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    ATCC hcov hku1 c n5
    Hcov Hku1 C N5, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hcov hku1 c n5  (ATCC)


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    ATCC hcov hku1 c n5
    Hcov Hku1 C N5, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hcov hku1 s  (ATCC)


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    ATCC hcov hku1 s
    A. Serum IgG binding titer against RBD (B.1.351). B. Comparison of serum neutralizing titers against VSV-SARS-CoV-2 Wu and B.1.351 at M1. Black horizontal line represents the mean, and the dashed line indicates no reduction. C. Frequency of Wu-specific and Wu+B.1.351 CR clonotypes in the RBD-pulldown. The total number of clonotypes is included at the center of each plot. D. Relative abundance of Wu+B.1.351 CR clonotypes in the RBD-pulldown. Relative abundance is calculated using the proteomics-based quantification of each antibody clonotype in serum. Mean values are represented as horizontal lines with the error bars indicating SD. E. Correlation between serum IgG binding titers to RBD (Wu) scaled by abundance of Wu+B.1.351 CR antibodies based on the proteomics data and serum IgG binding titers to RBD (B.1.351). F. Frequency of Wu-specific and Wu+B.1.351 CR antibody clonotypes that are S–RBD or iso– RBD. The error bars indicate SD. G. Serum IgG binding titer against <t>HKU1</t> S. H. Summary of <t>HKU1</t> <t>S</t> cross-reactive antibodies. For A and G , all samples run in technical duplicate, and the error bars indicate SD.
    Hcov Hku1 S, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Characterization of SARS-CoV-2 Convalescent Patients’ Serological Repertoire Reveals High Prevalence of Iso–RBD Antibodies"

    Article Title: Characterization of SARS-CoV-2 Convalescent Patients’ Serological Repertoire Reveals High Prevalence of Iso–RBD Antibodies

    Journal: bioRxiv

    doi: 10.1101/2023.09.08.556349

    A. Serum IgG binding titer against RBD (B.1.351). B. Comparison of serum neutralizing titers against VSV-SARS-CoV-2 Wu and B.1.351 at M1. Black horizontal line represents the mean, and the dashed line indicates no reduction. C. Frequency of Wu-specific and Wu+B.1.351 CR clonotypes in the RBD-pulldown. The total number of clonotypes is included at the center of each plot. D. Relative abundance of Wu+B.1.351 CR clonotypes in the RBD-pulldown. Relative abundance is calculated using the proteomics-based quantification of each antibody clonotype in serum. Mean values are represented as horizontal lines with the error bars indicating SD. E. Correlation between serum IgG binding titers to RBD (Wu) scaled by abundance of Wu+B.1.351 CR antibodies based on the proteomics data and serum IgG binding titers to RBD (B.1.351). F. Frequency of Wu-specific and Wu+B.1.351 CR antibody clonotypes that are S–RBD or iso– RBD. The error bars indicate SD. G. Serum IgG binding titer against HKU1 S. H. Summary of HKU1 S cross-reactive antibodies. For A and G , all samples run in technical duplicate, and the error bars indicate SD.
    Figure Legend Snippet: A. Serum IgG binding titer against RBD (B.1.351). B. Comparison of serum neutralizing titers against VSV-SARS-CoV-2 Wu and B.1.351 at M1. Black horizontal line represents the mean, and the dashed line indicates no reduction. C. Frequency of Wu-specific and Wu+B.1.351 CR clonotypes in the RBD-pulldown. The total number of clonotypes is included at the center of each plot. D. Relative abundance of Wu+B.1.351 CR clonotypes in the RBD-pulldown. Relative abundance is calculated using the proteomics-based quantification of each antibody clonotype in serum. Mean values are represented as horizontal lines with the error bars indicating SD. E. Correlation between serum IgG binding titers to RBD (Wu) scaled by abundance of Wu+B.1.351 CR antibodies based on the proteomics data and serum IgG binding titers to RBD (B.1.351). F. Frequency of Wu-specific and Wu+B.1.351 CR antibody clonotypes that are S–RBD or iso– RBD. The error bars indicate SD. G. Serum IgG binding titer against HKU1 S. H. Summary of HKU1 S cross-reactive antibodies. For A and G , all samples run in technical duplicate, and the error bars indicate SD.

    Techniques Used: Binding Assay, Comparison

    A. Binding specificity and affinity measured by ELISA. EC 50 values for each mAb to prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1.351), RBD (B.1.351), prefusion-stabilized S (B.1.1.529), and HKU1 S are shown. All assays were performed in triplicate. B-C. Neutralization activity of mAbs against pseudotyped VSV SARS-CoV-2 (Wu) ( B ) and VSV-SARS-CoV-2 (B.1.351) ( C ). CR3022, a non-neutralizing mAb was used as a negative control. The error bars indicate SD. D-F. Antibody-dependent cellular phagocytosis (ADCP) ( D ), FcγRIIIa activation (ADCC) ( E ), and antibody-dependent complement deposition (ADCD) ( F ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. REGN10987 IgG2σ was used as a null Fc-mediated effector function control. The dotted lines indicate the no antibody control.
    Figure Legend Snippet: A. Binding specificity and affinity measured by ELISA. EC 50 values for each mAb to prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1.351), RBD (B.1.351), prefusion-stabilized S (B.1.1.529), and HKU1 S are shown. All assays were performed in triplicate. B-C. Neutralization activity of mAbs against pseudotyped VSV SARS-CoV-2 (Wu) ( B ) and VSV-SARS-CoV-2 (B.1.351) ( C ). CR3022, a non-neutralizing mAb was used as a negative control. The error bars indicate SD. D-F. Antibody-dependent cellular phagocytosis (ADCP) ( D ), FcγRIIIa activation (ADCC) ( E ), and antibody-dependent complement deposition (ADCD) ( F ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. REGN10987 IgG2σ was used as a null Fc-mediated effector function control. The dotted lines indicate the no antibody control.

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Activity Assay, Negative Control, Activation Assay

    Antibody-dependent cellular phagocytosis ( A ), FcγRIIIa activation ( B ), and antibody-dependent complement deposition ( C ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. The dotted line indicates the no antibody control, and REGN10987 IgG2σ was used as a null Fc-mediated effector function control.
    Figure Legend Snippet: Antibody-dependent cellular phagocytosis ( A ), FcγRIIIa activation ( B ), and antibody-dependent complement deposition ( C ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. The dotted line indicates the no antibody control, and REGN10987 IgG2σ was used as a null Fc-mediated effector function control.

    Techniques Used: Activation Assay, Activity Assay

    hku1  (ATCC)


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    ATCC hku1
    Hku1, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hcov hku1 c n5  (ATCC)


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    86
    ATCC hku1
    Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and <t>-HKU1</t> (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.
    Hku1, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC hcov hku1
    Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and <t>-HKU1</t> (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.
    Hcov Hku1, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    ATCC hcov hku1 c n5
    Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and <t>-HKU1</t> (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.
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    86
    ATCC hcov hku1 s
    A. Serum IgG binding titer against RBD (B.1.351). B. Comparison of serum neutralizing titers against VSV-SARS-CoV-2 Wu and B.1.351 at M1. Black horizontal line represents the mean, and the dashed line indicates no reduction. C. Frequency of Wu-specific and Wu+B.1.351 CR clonotypes in the RBD-pulldown. The total number of clonotypes is included at the center of each plot. D. Relative abundance of Wu+B.1.351 CR clonotypes in the RBD-pulldown. Relative abundance is calculated using the proteomics-based quantification of each antibody clonotype in serum. Mean values are represented as horizontal lines with the error bars indicating SD. E. Correlation between serum IgG binding titers to RBD (Wu) scaled by abundance of Wu+B.1.351 CR antibodies based on the proteomics data and serum IgG binding titers to RBD (B.1.351). F. Frequency of Wu-specific and Wu+B.1.351 CR antibody clonotypes that are S–RBD or iso– RBD. The error bars indicate SD. G. Serum IgG binding titer against <t>HKU1</t> S. H. Summary of <t>HKU1</t> <t>S</t> cross-reactive antibodies. For A and G , all samples run in technical duplicate, and the error bars indicate SD.
    Hcov Hku1 S, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and -HKU1 (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.

    Journal: bioRxiv

    Article Title: Characterization of spike processing and entry mechanisms of seasonal human coronaviruses NL63, 229E and HKU1

    doi: 10.1101/2024.04.12.589332

    Figure Lengend Snippet: Infectivity of pseudoviruses bearing wild type (WT), monobasic furin cleavage site (mFCS) and deleted furin cleavage site (ΔFCS) spike proteins of HCoV-NL63 (A), -229E (C), and -HKU1 (E) in 293T-ACE2, 293T-ACE2-APN, and 293T-TMPRSS2. Dotted line indicates background level (1 × 10 4 RLU/ml). Data are shown as means and standard deviations from three independent experiments. The tests for two-group comparison (mutant spike vs WT spike) were analyzed using GraphPad Prism software. Asterisk* denotes significance: ****: p ≤ 0.0001 . Western blotting of HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 pseudoviruses (B, D and F) bearing indicated spike proteins. HCoV-NL63 (B), -229E (D), -HKU1 (F) and SARS-CoV-2 spike proteins were probed using anti-NL63-S1 mouse mAb (9D3F3), anti-229E-S1 mouse mAb (24C9F7), anti-HKU1-S1 rabbit polyclonal sera and rabbit pAb (Sino Biological, 40589-T62) respectively, Full length S0 and furin processed S1 subunits were indicated using arrows. HIV p24 probed using mouse hybridoma (183-H12-5) is shown for pseudovirus control.

    Article Snippet: Pseudoviruses bearing the desired spike glycoprotein of HCoV-NL63, -229E, -HKU1 and SARS-CoV-2 and carrying a firefly luciferase (FLuc) reporter gene were produced by cotransfecting HEK-293T (ATCC, Cat no: CRL-11268) cells with 5 μg of pCMVΔR8.2, 5 μg of pHR’CMVLuc and 0.5 μg of pVRC spike or its corresponding mutant expression plasmids.

    Techniques: Infection, Comparison, Mutagenesis, Software, Western Blot

    (A) Chloroquine sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (B) Camostat mesylate sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (C) Chloroquine sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (D) Ammonium chloride sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (E) Chloroquine sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2 cells. (F) Camostat mesylate sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2-TMPRSS2 cells respectively. For all experiments, cells were pretreated with indicated inhibitor for 2 h prior to pseudovirus infection in the presence of inhibitor for a period of 48 h. X-axis indicates log concentration of inhibitor. Y axis indicates percentage inhibition compared to pseudovirus infection without inhibitor treatment. Results shown are representative of three independent experiments.

    Journal: bioRxiv

    Article Title: Characterization of spike processing and entry mechanisms of seasonal human coronaviruses NL63, 229E and HKU1

    doi: 10.1101/2024.04.12.589332

    Figure Lengend Snippet: (A) Chloroquine sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (B) Camostat mesylate sensitivity of HCoV-NL63 and SARS-CoV-2 pseudoviruses in 293T-ACE2 and 293T-ACE2-TMPRSS2 cells. (C) Chloroquine sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (D) Ammonium chloride sensitivity of HCoV-229E pseudoviruses in 293T-ACE2-APN cells. (E) Chloroquine sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2 cells. (F) Camostat mesylate sensitivity of HCoV-HKU1 and SARS-CoV-2 pseudoviruses in 293T-TMPRSS2 and 293T-ACE2-TMPRSS2 cells respectively. For all experiments, cells were pretreated with indicated inhibitor for 2 h prior to pseudovirus infection in the presence of inhibitor for a period of 48 h. X-axis indicates log concentration of inhibitor. Y axis indicates percentage inhibition compared to pseudovirus infection without inhibitor treatment. Results shown are representative of three independent experiments.

    Article Snippet: Pseudoviruses bearing the desired spike glycoprotein of HCoV-NL63, -229E, -HKU1 and SARS-CoV-2 and carrying a firefly luciferase (FLuc) reporter gene were produced by cotransfecting HEK-293T (ATCC, Cat no: CRL-11268) cells with 5 μg of pCMVΔR8.2, 5 μg of pHR’CMVLuc and 0.5 μg of pVRC spike or its corresponding mutant expression plasmids.

    Techniques: Infection, Concentration Assay, Inhibition

    (A) Cellular TMPRSS2 is a type II transmembrane protease comprising N-terminal transmembrane (TM) domain, Low Density Lipoprotein Receptor domain A (LDLRA), Class A Scavenger Receptor Cysteine-Rich (SRCR) domain and a C-terminal trypsin-like serine protease (SP) domain with a canonical H296-D345-S441 catalytic triad. TMPRSS2 with an N-terminal Flag tag, LDLRA domain of TMPRSS2 with an N-terminal Flag tag, LDLRA and SRCR domains of TMPRSS2 with an N-terminal Flag tag, and TMPRSS2 proteins with alanine substitutions in the indicated active site residues of catalytic triad are shown. (B) Western Blotting of Flag-tagged TMPRSS2 proteins in 293T cells transfected with indicated plasmids. Lane 1 is Marker, Lane 2 showed TMPRSS2 expressed as full-length form (∼54KD) and cleavage products (∼20-25KD, 37-40KD) due to autoactivation. Lane 3-5 showed TMPRSS2 proteins lacking protease catalytic acitivity expressed mainly as full-length forms (∼54KD). Lanes 6 and show LDLRA and LDLRA-SRCR domains expressed as 25KD and 37KD forms. Lane shows Flag-tagged ACE2 (∼110KD) as a control. (C) Infectivity of VSV pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of VSV pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue, and green bars respectively (D) Infectivity of SARSCoV-2 pseudovirses in 293T-ACE2 cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of SARS-CoV-2 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively (E) Infectivity of HCoV-HKU1 pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of HCoV-HKU1 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively. The tests for two-group comparison (infectivity without inhibitor vs infectivity with inhibitor) were analyzed using GraphPad Prism software. Asterisk* denotes significance: *: p ≤ 0.05; **: p ≤ 0.01; ****: p ≤ 0.0001 .

    Journal: bioRxiv

    Article Title: Characterization of spike processing and entry mechanisms of seasonal human coronaviruses NL63, 229E and HKU1

    doi: 10.1101/2024.04.12.589332

    Figure Lengend Snippet: (A) Cellular TMPRSS2 is a type II transmembrane protease comprising N-terminal transmembrane (TM) domain, Low Density Lipoprotein Receptor domain A (LDLRA), Class A Scavenger Receptor Cysteine-Rich (SRCR) domain and a C-terminal trypsin-like serine protease (SP) domain with a canonical H296-D345-S441 catalytic triad. TMPRSS2 with an N-terminal Flag tag, LDLRA domain of TMPRSS2 with an N-terminal Flag tag, LDLRA and SRCR domains of TMPRSS2 with an N-terminal Flag tag, and TMPRSS2 proteins with alanine substitutions in the indicated active site residues of catalytic triad are shown. (B) Western Blotting of Flag-tagged TMPRSS2 proteins in 293T cells transfected with indicated plasmids. Lane 1 is Marker, Lane 2 showed TMPRSS2 expressed as full-length form (∼54KD) and cleavage products (∼20-25KD, 37-40KD) due to autoactivation. Lane 3-5 showed TMPRSS2 proteins lacking protease catalytic acitivity expressed mainly as full-length forms (∼54KD). Lanes 6 and show LDLRA and LDLRA-SRCR domains expressed as 25KD and 37KD forms. Lane shows Flag-tagged ACE2 (∼110KD) as a control. (C) Infectivity of VSV pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of VSV pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue, and green bars respectively (D) Infectivity of SARSCoV-2 pseudovirses in 293T-ACE2 cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of SARS-CoV-2 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively (E) Infectivity of HCoV-HKU1 pseudovirses in 293T cells transfected with indicated TMPRSS2 plasmids (black bars). Infectivity of HCoV-HKU1 pseudoviruses in the presence of camostat, chloroquine and NH 4 Cl are indicated in red, blue and green bars respectively. The tests for two-group comparison (infectivity without inhibitor vs infectivity with inhibitor) were analyzed using GraphPad Prism software. Asterisk* denotes significance: *: p ≤ 0.05; **: p ≤ 0.01; ****: p ≤ 0.0001 .

    Article Snippet: Pseudoviruses bearing the desired spike glycoprotein of HCoV-NL63, -229E, -HKU1 and SARS-CoV-2 and carrying a firefly luciferase (FLuc) reporter gene were produced by cotransfecting HEK-293T (ATCC, Cat no: CRL-11268) cells with 5 μg of pCMVΔR8.2, 5 μg of pHR’CMVLuc and 0.5 μg of pVRC spike or its corresponding mutant expression plasmids.

    Techniques: FLAG-tag, Western Blot, Transfection, Marker, Infection, Comparison, Software

    (A) Western blotting of HCoV-HKU1 spike in Anti-Flag Ab pull downs immunoprecipitated by indicated TMPRSS2 proteins with a Flag tag at the N-terminus. HCoV-HKU1 spike is probed using polyclonal serum. (B) Sample input employed for immunoprecipitation. TMPRSS2 proteins Flag tagged in the N-terminus are probed using anti-Flag antibody. Β-actin is loading control. (C) HCoV-HKU1 spike is expressed as both full length S0 and furin processed S1 forms. HCoV-HKU1-ΔFCS, which expresses only the full length S0 form is shown for size comparison.

    Journal: bioRxiv

    Article Title: Characterization of spike processing and entry mechanisms of seasonal human coronaviruses NL63, 229E and HKU1

    doi: 10.1101/2024.04.12.589332

    Figure Lengend Snippet: (A) Western blotting of HCoV-HKU1 spike in Anti-Flag Ab pull downs immunoprecipitated by indicated TMPRSS2 proteins with a Flag tag at the N-terminus. HCoV-HKU1 spike is probed using polyclonal serum. (B) Sample input employed for immunoprecipitation. TMPRSS2 proteins Flag tagged in the N-terminus are probed using anti-Flag antibody. Β-actin is loading control. (C) HCoV-HKU1 spike is expressed as both full length S0 and furin processed S1 forms. HCoV-HKU1-ΔFCS, which expresses only the full length S0 form is shown for size comparison.

    Article Snippet: Pseudoviruses bearing the desired spike glycoprotein of HCoV-NL63, -229E, -HKU1 and SARS-CoV-2 and carrying a firefly luciferase (FLuc) reporter gene were produced by cotransfecting HEK-293T (ATCC, Cat no: CRL-11268) cells with 5 μg of pCMVΔR8.2, 5 μg of pHR’CMVLuc and 0.5 μg of pVRC spike or its corresponding mutant expression plasmids.

    Techniques: Western Blot, Immunoprecipitation, FLAG-tag, Comparison

    A. Serum IgG binding titer against RBD (B.1.351). B. Comparison of serum neutralizing titers against VSV-SARS-CoV-2 Wu and B.1.351 at M1. Black horizontal line represents the mean, and the dashed line indicates no reduction. C. Frequency of Wu-specific and Wu+B.1.351 CR clonotypes in the RBD-pulldown. The total number of clonotypes is included at the center of each plot. D. Relative abundance of Wu+B.1.351 CR clonotypes in the RBD-pulldown. Relative abundance is calculated using the proteomics-based quantification of each antibody clonotype in serum. Mean values are represented as horizontal lines with the error bars indicating SD. E. Correlation between serum IgG binding titers to RBD (Wu) scaled by abundance of Wu+B.1.351 CR antibodies based on the proteomics data and serum IgG binding titers to RBD (B.1.351). F. Frequency of Wu-specific and Wu+B.1.351 CR antibody clonotypes that are S–RBD or iso– RBD. The error bars indicate SD. G. Serum IgG binding titer against HKU1 S. H. Summary of HKU1 S cross-reactive antibodies. For A and G , all samples run in technical duplicate, and the error bars indicate SD.

    Journal: bioRxiv

    Article Title: Characterization of SARS-CoV-2 Convalescent Patients’ Serological Repertoire Reveals High Prevalence of Iso–RBD Antibodies

    doi: 10.1101/2023.09.08.556349

    Figure Lengend Snippet: A. Serum IgG binding titer against RBD (B.1.351). B. Comparison of serum neutralizing titers against VSV-SARS-CoV-2 Wu and B.1.351 at M1. Black horizontal line represents the mean, and the dashed line indicates no reduction. C. Frequency of Wu-specific and Wu+B.1.351 CR clonotypes in the RBD-pulldown. The total number of clonotypes is included at the center of each plot. D. Relative abundance of Wu+B.1.351 CR clonotypes in the RBD-pulldown. Relative abundance is calculated using the proteomics-based quantification of each antibody clonotype in serum. Mean values are represented as horizontal lines with the error bars indicating SD. E. Correlation between serum IgG binding titers to RBD (Wu) scaled by abundance of Wu+B.1.351 CR antibodies based on the proteomics data and serum IgG binding titers to RBD (B.1.351). F. Frequency of Wu-specific and Wu+B.1.351 CR antibody clonotypes that are S–RBD or iso– RBD. The error bars indicate SD. G. Serum IgG binding titer against HKU1 S. H. Summary of HKU1 S cross-reactive antibodies. For A and G , all samples run in technical duplicate, and the error bars indicate SD.

    Article Snippet: Briefly, 500,000 1 μm yellow-green fluorescent microspheres (Thermo, F8813) were covalently conjugated with recombinant SARS-CoV-2 S, SARS-CoV-2 RBD, SARS-CoV-2 S (B.1.351), SARS-CoV-2 RBD (B.1.351), and hCoV HKU1 S and incubated for 3 hr with diluted recombinant mAbs (3-fold dilution, 5 μg/mL to 6.8 ng/mL) and 20,000 cells from the human monocytic THP-1 cell line (ATCC, TIB-202).

    Techniques: Binding Assay, Comparison

    A. Binding specificity and affinity measured by ELISA. EC 50 values for each mAb to prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1.351), RBD (B.1.351), prefusion-stabilized S (B.1.1.529), and HKU1 S are shown. All assays were performed in triplicate. B-C. Neutralization activity of mAbs against pseudotyped VSV SARS-CoV-2 (Wu) ( B ) and VSV-SARS-CoV-2 (B.1.351) ( C ). CR3022, a non-neutralizing mAb was used as a negative control. The error bars indicate SD. D-F. Antibody-dependent cellular phagocytosis (ADCP) ( D ), FcγRIIIa activation (ADCC) ( E ), and antibody-dependent complement deposition (ADCD) ( F ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. REGN10987 IgG2σ was used as a null Fc-mediated effector function control. The dotted lines indicate the no antibody control.

    Journal: bioRxiv

    Article Title: Characterization of SARS-CoV-2 Convalescent Patients’ Serological Repertoire Reveals High Prevalence of Iso–RBD Antibodies

    doi: 10.1101/2023.09.08.556349

    Figure Lengend Snippet: A. Binding specificity and affinity measured by ELISA. EC 50 values for each mAb to prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1.351), RBD (B.1.351), prefusion-stabilized S (B.1.1.529), and HKU1 S are shown. All assays were performed in triplicate. B-C. Neutralization activity of mAbs against pseudotyped VSV SARS-CoV-2 (Wu) ( B ) and VSV-SARS-CoV-2 (B.1.351) ( C ). CR3022, a non-neutralizing mAb was used as a negative control. The error bars indicate SD. D-F. Antibody-dependent cellular phagocytosis (ADCP) ( D ), FcγRIIIa activation (ADCC) ( E ), and antibody-dependent complement deposition (ADCD) ( F ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. REGN10987 IgG2σ was used as a null Fc-mediated effector function control. The dotted lines indicate the no antibody control.

    Article Snippet: Briefly, 500,000 1 μm yellow-green fluorescent microspheres (Thermo, F8813) were covalently conjugated with recombinant SARS-CoV-2 S, SARS-CoV-2 RBD, SARS-CoV-2 S (B.1.351), SARS-CoV-2 RBD (B.1.351), and hCoV HKU1 S and incubated for 3 hr with diluted recombinant mAbs (3-fold dilution, 5 μg/mL to 6.8 ng/mL) and 20,000 cells from the human monocytic THP-1 cell line (ATCC, TIB-202).

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Activity Assay, Negative Control, Activation Assay

    Antibody-dependent cellular phagocytosis ( A ), FcγRIIIa activation ( B ), and antibody-dependent complement deposition ( C ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. The dotted line indicates the no antibody control, and REGN10987 IgG2σ was used as a null Fc-mediated effector function control.

    Journal: bioRxiv

    Article Title: Characterization of SARS-CoV-2 Convalescent Patients’ Serological Repertoire Reveals High Prevalence of Iso–RBD Antibodies

    doi: 10.1101/2023.09.08.556349

    Figure Lengend Snippet: Antibody-dependent cellular phagocytosis ( A ), FcγRIIIa activation ( B ), and antibody-dependent complement deposition ( C ) activity of mAbs against SARS-CoV-2 prefusion-stabilized S (Wu), RBD (Wu), prefusion-stabilized S (B.1351), and HKU1 S. The dotted line indicates the no antibody control, and REGN10987 IgG2σ was used as a null Fc-mediated effector function control.

    Article Snippet: Briefly, 500,000 1 μm yellow-green fluorescent microspheres (Thermo, F8813) were covalently conjugated with recombinant SARS-CoV-2 S, SARS-CoV-2 RBD, SARS-CoV-2 S (B.1.351), SARS-CoV-2 RBD (B.1.351), and hCoV HKU1 S and incubated for 3 hr with diluted recombinant mAbs (3-fold dilution, 5 μg/mL to 6.8 ng/mL) and 20,000 cells from the human monocytic THP-1 cell line (ATCC, TIB-202).

    Techniques: Activation Assay, Activity Assay