anti s2  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike Neutralizing Antibody Mouse Mab
    Description:
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified Recombinant SARS CoV 2 2019 nCoV Spike S1 mFc Protein Catalog 40591 V05H1 YP 009724390 1 Met1 Arg685 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    Catalog Number:
    40591-MM43
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    2019 nCoV
    Applications:
    ELISA,Neutralization
    Immunogen:
    Recombinant SARS-CoV-2 (2019-nCoV) Spike S1-mFc Protein (Catalog#40591-V05H1)
    Product Aliases:
    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-NCP-CoV RBD Antibody, Anti-NCP-CoV s1 Antibody, Anti-NCP-CoV s2 Antibody, Anti-NCP-CoV Spike Antibody, Anti-novel coronavirus RBD Antibody, Anti-novel coronavirus s1 Antibody, Anti-novel coronavirus s2 Antibody, Anti-novel coronavirus spike Antibody, Anti-RBD Antibody, Anti-S1 Antibody, Anti-S2 Antibody, Anti-Spike RBD Antibody
    Antibody Type:
    MAb
    Host:
    Mouse
    Isotype:
    Mouse IgG1
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    Structured Review

    Sino Biological anti s2
    N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using <t>anti-S2</t> Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified Recombinant SARS CoV 2 2019 nCoV Spike S1 mFc Protein Catalog 40591 V05H1 YP 009724390 1 Met1 Arg685 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    https://www.bioz.com/result/anti s2/product/Sino Biological
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti s2 - by Bioz Stars, 2021-06
    99/100 stars

    Images

    1) Product Images from "Inhibition of SARS-CoV-2 viral entry in vitro upon blocking N- and O-glycan elaboration"

    Article Title: Inhibition of SARS-CoV-2 viral entry in vitro upon blocking N- and O-glycan elaboration

    Journal: bioRxiv

    doi: 10.1101/2020.10.15.339838

    N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P
    Figure Legend Snippet: N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P

    Techniques Used: Modification, Expressing, Mutagenesis, Produced, Generated, Titration, Infection, Western Blot

    2) Product Images from "Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration"

    Article Title: Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration

    Journal: eLife

    doi: 10.7554/eLife.61552

    N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. ( A ) Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] - and [N] - 293 T cells. All nine viruses were applied at equal titer to stable 293T/ACE2. ( B–C ) O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. ( D ) Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. ( E ) Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] - 293T products due to truncation of glycan biosynthesis. ( F ) Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] - 293Ts is almost fully proteolyzed during viral production (red arrowhead). *p
    Figure Legend Snippet: N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. ( A ) Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] - and [N] - 293 T cells. All nine viruses were applied at equal titer to stable 293T/ACE2. ( B–C ) O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. ( D ) Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. ( E ) Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] - 293T products due to truncation of glycan biosynthesis. ( F ) Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] - 293Ts is almost fully proteolyzed during viral production (red arrowhead). *p

    Techniques Used: Modification, Expressing, Mutagenesis, Produced, Generated, Titration, Infection, Western Blot

    3) Product Images from "Inhibition of SARS-CoV-2 viral entry in vitro upon blocking N- and O-glycan elaboration"

    Article Title: Inhibition of SARS-CoV-2 viral entry in vitro upon blocking N- and O-glycan elaboration

    Journal: bioRxiv

    doi: 10.1101/2020.10.15.339838

    N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P
    Figure Legend Snippet: N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P

    Techniques Used: Modification, Expressing, Mutagenesis, Produced, Generated, Titration, Infection, Western Blot

    Related Articles

    Western Blot:

    Article Title: Inhibition of SARS-CoV-2 viral entry in vitro upon blocking N- and O-glycan elaboration
    Article Snippet: Identity of expressed protein and also viral Spike was determined using western blotting with anti-Fc (Jackson), anti-RBD (Sino Biologicals), anti-S2 (Sino Biologicals) and anti-ACE2 (R & D Systems) pAbs. .. Identity of expressed protein and also viral Spike was determined using western blotting with anti-Fc (Jackson), anti-RBD (Sino Biologicals), anti-S2 (Sino Biologicals) and anti-ACE2 (R & D Systems) pAbs. .. Anti-FLAG mAb L5 was also used for western blotting of Spike-mutant virus as it carried a C-terminal FLAG tag.

    Article Title: Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration
    Article Snippet: Final protein concentration was determined in ELISA format by using a calibrant Fc-protein of known concentration that was verified to be > 95% pure based on silver stain analysis. .. Identity of expressed protein and also viral Spike was determined using western blotting with anti-Fc (Jackson), anti-RBD (Sino Biologicals), anti-S2 (Sino Biologicals) and anti-ACE2 (R and D Systems) pAbs. .. Anti-FLAG mAb L5 was also used for western blotting of Spike-mutant virus as it carried a C-terminal FLAG tag.

    Mouse Assay:

    Article Title: Virus‐Free and Live‐Cell Visualizing SARS‐CoV‐2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors, Virus‐Free and Live‐Cell Visualizing SARS‐CoV‐2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors
    Article Snippet: .. Generation and Production of Antibodies against SARS‐CoV‐2 S Balb/c mice were intraperitoneal immunized with 5 µg of SARS‐CoV2‐RBD (expression in this study, n = 5), SARS‐CoV2‐S1 (Sino Biological, 40591‐V08H, n = 3), and SARS‐CoV2‐S2 (Sino Biological, 40590‐V08B, n = 3), respectively. ..

    Expressing:

    Article Title: Virus‐Free and Live‐Cell Visualizing SARS‐CoV‐2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors, Virus‐Free and Live‐Cell Visualizing SARS‐CoV‐2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors
    Article Snippet: .. Generation and Production of Antibodies against SARS‐CoV‐2 S Balb/c mice were intraperitoneal immunized with 5 µg of SARS‐CoV2‐RBD (expression in this study, n = 5), SARS‐CoV2‐S1 (Sino Biological, 40591‐V08H, n = 3), and SARS‐CoV2‐S2 (Sino Biological, 40590‐V08B, n = 3), respectively. ..

    Infection:

    Article Title: A single dose of recombinant VSV-ΔG-spike vaccine provides protection against SARS-CoV-2 challenge
    Article Snippet: The hamster model is a robust and reproducible model as evident by the dose-dependent response in body weight to different SARS-CoV-2 infection doses. .. The hamster model is a robust and reproducible model as evident by the dose-dependent response in body weight to different SARS-CoV-2 infection doses. ..

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  • 99
    Sino Biological anti s2
    N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using <t>anti-S2</t> Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P
    Anti S2, supplied by Sino Biological, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti s2/product/Sino Biological
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti s2 - by Bioz Stars, 2021-06
    99/100 stars
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    95
    Sino Biological s2 subunit
    RXXR motifs located at the border between S1 and S2 are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein (MERS-S). A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the <t>S2</t> subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.
    S2 Subunit, 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/s2 subunit/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    s2 subunit - by Bioz Stars, 2021-06
    95/100 stars
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    98
    Sino Biological sars cov 2 2019 ncov spike s2 antibody rabbit pab
    Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin <t>SARS-CoV-2pp</t> were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the <t>SARS-CoV-2</t> S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.
    Sars Cov 2 2019 Ncov Spike S2 Antibody Rabbit Pab, supplied by Sino Biological, used in various techniques. Bioz Stars score: 98/100, 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 s2 antibody rabbit pab/product/Sino Biological
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike s2 antibody rabbit pab - by Bioz Stars, 2021-06
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    N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P

    Journal: bioRxiv

    Article Title: Inhibition of SARS-CoV-2 viral entry in vitro upon blocking N- and O-glycan elaboration

    doi: 10.1101/2020.10.15.339838

    Figure Lengend Snippet: N-glycan modification of SARS-CoV-2 pseudovirus abolishes entry into 293T/ACE2 cells. A . Pseudovirus expressing VSVG envelope protein, Spike-WT and Spike-mutant were produced in wild-type, [O] − and [N] − 293T cells. All 9 viruses were applied at equal titer to stable 293T/ACE2. B - C . O-glycan truncation of Spike partially reduced viral entry. N-glycan truncation abolished viral entry. In order to combine data from multiple viral preparations and independent runs in a single plot, all data were normalized by setting DsRed signal produced by virus generated in wild-type 293T to 10,000 normalized MFI or 100% normalized DsRed positive value. D . Viral titration study performed with Spike-mutant virus shows complete loss of viral infection over a wide range. E . Western blot of Spike protein using anti-S2 Ab shows reduced proteolysis of Spike-mut compared to Spike-WT. The full Spike protein and free S2-subunit resulting from S1-S2 cleavage is indicated. Molecular mass is reduced in [N] − 293T products due to truncation of glycan biosynthesis. F . Anti-FLAG Ab binds the C-terminus of Spike-mutant. Spike produced in [N] − 293Ts is almost fully proteolyzed during viral production (red arrowhead). * P

    Article Snippet: Identity of expressed protein and also viral Spike was determined using western blotting with anti-Fc (Jackson), anti-RBD (Sino Biologicals), anti-S2 (Sino Biologicals) and anti-ACE2 (R & D Systems) pAbs.

    Techniques: Modification, Expressing, Mutagenesis, Produced, Generated, Titration, Infection, Western Blot

    RXXR motifs located at the border between S1 and S2 are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein (MERS-S). A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.

    Journal: The Journal of Infectious Diseases

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

    doi: 10.1093/infdis/jiu407

    Figure Lengend Snippet: RXXR motifs located at the border between S1 and S2 are required for efficient processing of the Middle East respiratory syndrome coronavirus spike protein (MERS-S). A , The domain organization of the MERS-S protein is schematically depicted. The MERS-S sequence at the border between the S1 and S2 subunits is shown. RXXR motifs, which constitute potential cleavage sites, are highlighted, and the predicted start of the S2 subunit is underlined. The mutations introduced into the potential cleavage sites in MERS-S are shown. B , 293T cells were transfected with expression plasmids coding for MERS-S wild type and the indicated MERS-S mutants equipped with a C-terminal V5 tag. Transfection of empty plasmid (pcDNA) served as negative control. Expression of S proteins in cell lysates was determined by Western blot, using a V5 tag–specific monoclonal antibody. Expression of β-actin in cell lysates was assessed as a loading control. The results shown are representative for at least 3 independent experiments. Abbreviations: CT, cytoplasmic tail; PCM, potential cleavage site mutant; RBD, receptor binding domain; SP, signal peptide; TM, transmembrane domain.

    Article Snippet: MERS-S expression was detected using a monoclonal antibody directed against the V5 tag (Invitrogen) or a polyclonal antibody directed against the S2 subunit of the MERS-S protein (Sino Biological).

    Techniques: Sequencing, Transfection, Expressing, Plasmid Preparation, Negative Control, Western Blot, Mutagenesis, Binding Assay

    The Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein (MERS-S) is cleaved in transfected and infected cells. 293T cells were transfected with a plasmid encoding the MERS-S protein or with empty plasmid (pcDNA). Vero B4 cells were either infected with MERS-CoV at a multiplicity of infection of 5 or mock infected. Subsequently, the cells were lysed and analyzed by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S. A β-actin antibody served as a loading control. Similar results were obtained in 2 separate experiments.

    Journal: The Journal of Infectious Diseases

    Article Title: Inhibition of Proprotein Convertases Abrogates Processing of the Middle Eastern Respiratory Syndrome Coronavirus Spike Protein in Infected Cells but Does Not Reduce Viral Infectivity

    doi: 10.1093/infdis/jiu407

    Figure Lengend Snippet: The Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein (MERS-S) is cleaved in transfected and infected cells. 293T cells were transfected with a plasmid encoding the MERS-S protein or with empty plasmid (pcDNA). Vero B4 cells were either infected with MERS-CoV at a multiplicity of infection of 5 or mock infected. Subsequently, the cells were lysed and analyzed by Western blot, using a polyclonal antibody directed against the S2 subunit of MERS-S. A β-actin antibody served as a loading control. Similar results were obtained in 2 separate experiments.

    Article Snippet: MERS-S expression was detected using a monoclonal antibody directed against the V5 tag (Invitrogen) or a polyclonal antibody directed against the S2 subunit of the MERS-S protein (Sino Biological).

    Techniques: Transfection, Infection, Plasmid Preparation, Western Blot

    Comparison of saliva and serum SARS-CoV-2 antigen-specific IgG responses by days post-symptom onset (DPSO). The trajectories of IgG responses (red solid lines) and confidence intervals (semitransparent background) were estimated using a LOESS curve. Dashed red lines indicate cutoff values for each antigen. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity; a.u., arbitrary units.

    Journal: Journal of Clinical Microbiology

    Article Title: COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva

    doi: 10.1128/JCM.02204-20

    Figure Lengend Snippet: Comparison of saliva and serum SARS-CoV-2 antigen-specific IgG responses by days post-symptom onset (DPSO). The trajectories of IgG responses (red solid lines) and confidence intervals (semitransparent background) were estimated using a LOESS curve. Dashed red lines indicate cutoff values for each antigen. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity; a.u., arbitrary units.

    Article Snippet: These included four SARS-CoV-2 receptor binding domain (RBD) proteins, one ectodomain (ECD) protein containing the S1 and S2 subunits of the spike protein, two S1 subunits, one S2 subunit, and two N proteins (see Table S1 in the supplemental material).

    Techniques: Binding Assay, Produced, Fluorescence

    Assay sensitivity and specificity for each SARS-CoV-2 antigen and antibody isotype using saliva (a) and serum (b). Samples collected from individuals with RT-PCR-confirmed prior SARS-CoV-2 infection are stratified by time since symptom onset. Darker shades of green indicate higher and darker shades of red indicate lower sensitivity and specificity. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells.

    Journal: Journal of Clinical Microbiology

    Article Title: COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva

    doi: 10.1128/JCM.02204-20

    Figure Lengend Snippet: Assay sensitivity and specificity for each SARS-CoV-2 antigen and antibody isotype using saliva (a) and serum (b). Samples collected from individuals with RT-PCR-confirmed prior SARS-CoV-2 infection are stratified by time since symptom onset. Darker shades of green indicate higher and darker shades of red indicate lower sensitivity and specificity. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells.

    Article Snippet: These included four SARS-CoV-2 receptor binding domain (RBD) proteins, one ectodomain (ECD) protein containing the S1 and S2 subunits of the spike protein, two S1 subunits, one S2 subunit, and two N proteins (see Table S1 in the supplemental material).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Infection, Binding Assay, Produced

    Correlation between matched SARS-CoV-2-specific IgG responses in saliva and serum ( n = 28). The Pearson correlation coefficient is provided for IgG responses to each antigen. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity.

    Journal: Journal of Clinical Microbiology

    Article Title: COVID-19 Serology at Population Scale: SARS-CoV-2-Specific Antibody Responses in Saliva

    doi: 10.1128/JCM.02204-20

    Figure Lengend Snippet: Correlation between matched SARS-CoV-2-specific IgG responses in saliva and serum ( n = 28). The Pearson correlation coefficient is provided for IgG responses to each antigen. Sino Biol., Sino Biological; NAC, Native Antigen Company; N, nucleocapsid protein; ECD, ectodomain; S1/S2, S1 or S2 subunit of the spike protein; RBD, receptor binding domain; (h), produced in human cells; (i), produced in insect cells; MFI, median fluorescence intensity.

    Article Snippet: These included four SARS-CoV-2 receptor binding domain (RBD) proteins, one ectodomain (ECD) protein containing the S1 and S2 subunits of the spike protein, two S1 subunits, one S2 subunit, and two N proteins (see Table S1 in the supplemental material).

    Techniques: Binding Assay, Produced, Fluorescence

    Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Produced, Transfection, Recombinant, Infection, Western Blot

    Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Infection, Produced, Concentration Assay, Cleavage Assay, Inhibition, Incubation, Recombinant

    Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Sequencing

    Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Infection, Luciferase, Activity Assay, Western Blot