sars cov 2 2019 ncov spike s2 ecd his recombinant protein covid 19 spike s2 research  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike S2 ECD His Recombinant Protein COVID 19 Spike S2 Research
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein S2 ECD YP 009724390 1 Ser686 Pro1213 was expressed with a polyhistidine tag at the C terminus
    Catalog Number:
    40590-v08b
    Price:
    None
    Category:
    recombinant protein
    Product Aliases:
    coronavirus spike Protein 2019-nCoV, cov spike Protein 2019-nCoV, ncov RBD Protein 2019-nCoV, ncov s1 Protein 2019-nCoV, ncov s2 Protein 2019-nCoV, ncov spike Protein 2019-nCoV, NCP-CoV RBD Protein 2019-nCoV, NCP-CoV s1 Protein 2019-nCoV, NCP-CoV s2 Protein 2019-nCoV, NCP-CoV Spike Protein 2019-nCoV, novel coronavirus RBD Protein 2019-nCoV, novel coronavirus s1 Protein 2019-nCoV, novel coronavirus s2 Protein 2019-nCoV, novel coronavirus spike Protein 2019-nCoV, RBD Protein 2019-nCoV, S1 Protein 2019-nCoV, S2 Protein 2019-nCoV, Spike RBD Protein 2019-nCoV
    Host:
    Baculovirus-Insect Cells
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    Structured Review

    Sino Biological sars cov 2 2019 ncov spike s2 ecd his recombinant protein covid 19 spike s2 research
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein S2 ECD YP 009724390 1 Ser686 Pro1213 was expressed with a polyhistidine tag at the C terminus
    https://www.bioz.com/result/sars cov 2 2019 ncov spike s2 ecd his recombinant protein covid 19 spike s2 research/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike s2 ecd his recombinant protein covid 19 spike s2 research - by Bioz Stars, 2021-03
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    Enzyme-linked Immunosorbent Assay:

    Article Title: Disease severity dictates SARS-CoV-2-specific neutralizing antibody responses in COVID-19
    Article Snippet: .. Enzyme-linked immunosorbent assay As previously described, 50 ng of SARS-CoV-2 S1 protein (Sino Biological, 40591-V08H) or SARS-CoV-2 RBD protein (Sino Biological, 40592-V08B) or SARS-CoV-2 S2 protein (Sino Biological, 40590-V08B) in 100 μl phosphate-buffered saline (PBS) per well was coated on ELISA plates (Costar, 42592) overnight at 4 °C. .. The ELISA plates were blocked for 1 h with 100 μl blocking buffer (5% fetal bovine serum (FBS) and 0.1% Tween 20 in PBS) and then incubated with diluted patient or healthy control sera in 100 μl blocking buffer for 1 h. After washing with PBST buffer (0.1% Tween 20 in PBS), the ELISA plates were incubated with anti-human IgG horseradish peroxidase (HRP) antibody (Bioss Biotech, 0297D) for 45 min, followed by PBST washing and addition of 3,3′,5,5′-tetramethylbenzidine (TMB) buffer (Beyotime, P0209).

    Construct:

    Article Title: Cold sensitivity of the SARS-CoV-2 spike ectodomain
    Article Snippet: Antibodies were incubated, washed and binding detected with goat anti-human-HRP (Jackson ImmunoResearch Laboratories, #109-035-098) and TMB substrate. .. Commercially obtained constructs of SARS-CoV-2 spike ectodomain (S1+S2 ECD, S2 ECD and RBD) (Sino Biological Inc cat# 40589-V08B1 and 40590-V08B respectively and RBD from Genescript cat# Z03483) were coated directly on 384-well plates at 2 µg/ml and incubated overnight at 4 °C. ..

    Article Title: A glycan cluster on the SARS-CoV-2 spike ectodomain is recognized by Fab-dimerized glycan-reactive antibodies
    Article Snippet: SARS-CoV-2 spike protein, and Man9 -V3 and Aglycone V3 peptides were captured via streptavidin on Nunc-absorb ELISA plates using PBS-based buffers and assay conditions as previously described ( , ). .. HIV-1 CH505TF SOSIP trimer and commercially-obtained constructs of SARS-CoV-2 spike ectodomain (S1+S2 ECD, S2 ECD and RBD) (Sino Biological Inc cat# 40589-V08B1 and 40590-V08B respectively and RBD from Genescript cat# Z03483) were captured using mouse anti-AVI-tag mAb (Avidity LLC, Aurora, CO). .. In brief, we coated 30ng of streptavidin in 15µl at 2µg/ml or 30 ng of mouse anti-AVI-tag mAb in 15µl at 2µg/ml per well of a 384-well Nunc-absorb ELISA plate, sealed and incubated overnight at 4°C.

    Incubation:

    Article Title: Cold sensitivity of the SARS-CoV-2 spike ectodomain
    Article Snippet: Antibodies were incubated, washed and binding detected with goat anti-human-HRP (Jackson ImmunoResearch Laboratories, #109-035-098) and TMB substrate. .. Commercially obtained constructs of SARS-CoV-2 spike ectodomain (S1+S2 ECD, S2 ECD and RBD) (Sino Biological Inc cat# 40589-V08B1 and 40590-V08B respectively and RBD from Genescript cat# Z03483) were coated directly on 384-well plates at 2 µg/ml and incubated overnight at 4 °C. ..

    Recombinant:

    Article Title: The receptor binding domain of SARS-CoV-2 spike is the key target of neutralizing antibody in human polyclonal sera
    Article Snippet: Recombinant ProteinsVaccinia Virus A33R Protein with C-Terminal Histidine Tag, was obtained through BEI Resources, NIAID, NIH: NR-2623. .. SARS-CoV-2 (2019-nCoV) Spike S1-His Recombinant Protein (40591-V08B1) and SARS-CoV-2 (2019-nCoV) Spike S2 ECD-His Recombinant Protein (40590-V08B) were obtained from Sino Biological. .. SARS-CoV-2 Spike protein (RBD, His Tag) (Z03483) was obtained though GenScript.

    Article Title: SARS-CoV-2 infection induces germinal center responses with robust stimulation of CD4 T follicular helper cells in rhesus macaques
    Article Snippet: .. BAMA for IgG and IgM antibodies to S1, S2 and N proteinsA customized BAMA was developed to simultaneously measure antibodies to the following recombinant SARS CoV-2 proteins (all from SinoBiologicals, Wayne, PA): S1 (#40591-V08H), S2 extracellular domain (#40590-V08B) and nucleocapsid (N; #40588-V08B). .. Briefly, proteins were dialyzed in PBS and conjugated to Bioplex Pro carboxylated magnetic beads (BioRad, Hercules, CA) as previously described ( ).

    Article Title: SARS-CoV-2 induces robust germinal center CD4 T follicular helper cell responses in rhesus macaques
    Article Snippet: .. Binding antibody multiplex assay (BAMA) for IgG and IgM antibodies to S1, S2 and N proteins A customized BAMA was developed to simultaneously measure antibodies to the following recombinant SARS-CoV-2 proteins (all from SinoBiologicals, Wayne, PA): S1 (#40591-V08H), S2 extracellular domain (#40590-V08B) and nucleocapsid (N; #40588-V08B). .. Briefly, proteins were dialyzed in PBS and conjugated to Bioplex Pro carboxylated magnetic beads (BioRad, Hercules, CA) .

    Binding Assay:

    Article Title: A rapidly adaptable biomaterial vaccine for SARS-CoV-2
    Article Snippet: Monophosphoryl lipid A (MPLA) derived from Salmonella minnesota R595 was purchased from Invivogen, USA (vac-mpla). .. SARS-CoV-2 antigens included spike protein subunits, S1 (40591-V08H), S2 (40590-V08B), and nucleocapsid (N) protein (40588-V08B) was purchased from Sino Biological, USA, and the receptor binding domain (RBD) protein (NR-52306) was purchased from BEI resources, USA. ..

    Article Title: SARS-CoV-2 induces robust germinal center CD4 T follicular helper cell responses in rhesus macaques
    Article Snippet: .. Binding antibody multiplex assay (BAMA) for IgG and IgM antibodies to S1, S2 and N proteins A customized BAMA was developed to simultaneously measure antibodies to the following recombinant SARS-CoV-2 proteins (all from SinoBiologicals, Wayne, PA): S1 (#40591-V08H), S2 extracellular domain (#40590-V08B) and nucleocapsid (N; #40588-V08B). .. Briefly, proteins were dialyzed in PBS and conjugated to Bioplex Pro carboxylated magnetic beads (BioRad, Hercules, CA) .

    Multiplex Assay:

    Article Title: SARS-CoV-2 induces robust germinal center CD4 T follicular helper cell responses in rhesus macaques
    Article Snippet: .. Binding antibody multiplex assay (BAMA) for IgG and IgM antibodies to S1, S2 and N proteins A customized BAMA was developed to simultaneously measure antibodies to the following recombinant SARS-CoV-2 proteins (all from SinoBiologicals, Wayne, PA): S1 (#40591-V08H), S2 extracellular domain (#40590-V08B) and nucleocapsid (N; #40588-V08B). .. Briefly, proteins were dialyzed in PBS and conjugated to Bioplex Pro carboxylated magnetic beads (BioRad, Hercules, CA) .

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    Sino Biological anti sars cov 2 d001
    RAPPID enables versatile monitoring of clinically relevant antibodies. a , Schematic overview of RAPPID sensors for the detection of anti-drug antibodies (ADAs). Use of a commercially available therapeutic antibody conjugated to both Gx-LB and Gx-SB enables binding of the ADA and formation of a luminescent ternary complex. b , Ratiometric sensor response curves of RAPPID sensors for anti-cetuximab, anti-adalimumab, and anti-infliximab, respectively. c , Schematic overview of RAPPID sensors for detection of therapeutic antibodies adalimumab and infliximab using target antigen TNFα fused to SB, and the anti-antibody conjugated to Gx-LB. d , Intensiometric detection of infliximab without calibrator luciferase. Luminescent measurements were performed at various intervals following addition of the NanoLuc substrate. e , Sensor response for adalimumab (left) and infliximab (right) detection in diluted plasma. The same sample was used for detection with a digital camera (top) and the plate reader (bottom). f , Schematic overview of RAPPID sensors for direct monitoring of anti-virus antibodies against the receptor binding domain (RBD) of <t>SARS-CoV-2</t> spike protein. LB and SB were expressed as fusion proteins with the RBD, enabling detection of all RBD-targeting antibodies. g , Ratiometric sensor response curves for neutralizing antibody 47D11 (left) 44 , and commercially available anti-spike <t>D001</t> (center) and D003 (right) 43 . Insets show sensor response at low target concentrations, which allowed calculation of the limit of detection, as indicated in the graphs. Experiments in b , d , e , g were performed in triplicate at sensor and calibrator concentrations as indicated in a , c , f , respectively. Reaction mixtures were prepared in buffer (PBS (pH 7.4), 0.1% (w/v) BSA) and incubated for 1 hr at room temperature before addition of NanoLuc substrate and recording of the emission spectra. The blue/green ratio was calculated by dividing bioluminescent emission at 458 nm by emission at 518 nm. The fold increase is indicated in each graph. Individual data points are represented as circles, while dashed lines connect mean values. Bars in histograms represent mean values.
    Anti Sars Cov 2 D001, 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
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    Average 94 stars, based on 1 article reviews
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    95
    Sino Biological sars cov 2
    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 <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, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 - by Bioz Stars, 2021-03
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    RAPPID enables versatile monitoring of clinically relevant antibodies. a , Schematic overview of RAPPID sensors for the detection of anti-drug antibodies (ADAs). Use of a commercially available therapeutic antibody conjugated to both Gx-LB and Gx-SB enables binding of the ADA and formation of a luminescent ternary complex. b , Ratiometric sensor response curves of RAPPID sensors for anti-cetuximab, anti-adalimumab, and anti-infliximab, respectively. c , Schematic overview of RAPPID sensors for detection of therapeutic antibodies adalimumab and infliximab using target antigen TNFα fused to SB, and the anti-antibody conjugated to Gx-LB. d , Intensiometric detection of infliximab without calibrator luciferase. Luminescent measurements were performed at various intervals following addition of the NanoLuc substrate. e , Sensor response for adalimumab (left) and infliximab (right) detection in diluted plasma. The same sample was used for detection with a digital camera (top) and the plate reader (bottom). f , Schematic overview of RAPPID sensors for direct monitoring of anti-virus antibodies against the receptor binding domain (RBD) of SARS-CoV-2 spike protein. LB and SB were expressed as fusion proteins with the RBD, enabling detection of all RBD-targeting antibodies. g , Ratiometric sensor response curves for neutralizing antibody 47D11 (left) 44 , and commercially available anti-spike D001 (center) and D003 (right) 43 . Insets show sensor response at low target concentrations, which allowed calculation of the limit of detection, as indicated in the graphs. Experiments in b , d , e , g were performed in triplicate at sensor and calibrator concentrations as indicated in a , c , f , respectively. Reaction mixtures were prepared in buffer (PBS (pH 7.4), 0.1% (w/v) BSA) and incubated for 1 hr at room temperature before addition of NanoLuc substrate and recording of the emission spectra. The blue/green ratio was calculated by dividing bioluminescent emission at 458 nm by emission at 518 nm. The fold increase is indicated in each graph. Individual data points are represented as circles, while dashed lines connect mean values. Bars in histograms represent mean values.

    Journal: bioRxiv

    Article Title: RAPPID: a platform of ratiometric bioluminescent sensors for homogeneous immunoassays

    doi: 10.1101/2020.10.31.363044

    Figure Lengend Snippet: RAPPID enables versatile monitoring of clinically relevant antibodies. a , Schematic overview of RAPPID sensors for the detection of anti-drug antibodies (ADAs). Use of a commercially available therapeutic antibody conjugated to both Gx-LB and Gx-SB enables binding of the ADA and formation of a luminescent ternary complex. b , Ratiometric sensor response curves of RAPPID sensors for anti-cetuximab, anti-adalimumab, and anti-infliximab, respectively. c , Schematic overview of RAPPID sensors for detection of therapeutic antibodies adalimumab and infliximab using target antigen TNFα fused to SB, and the anti-antibody conjugated to Gx-LB. d , Intensiometric detection of infliximab without calibrator luciferase. Luminescent measurements were performed at various intervals following addition of the NanoLuc substrate. e , Sensor response for adalimumab (left) and infliximab (right) detection in diluted plasma. The same sample was used for detection with a digital camera (top) and the plate reader (bottom). f , Schematic overview of RAPPID sensors for direct monitoring of anti-virus antibodies against the receptor binding domain (RBD) of SARS-CoV-2 spike protein. LB and SB were expressed as fusion proteins with the RBD, enabling detection of all RBD-targeting antibodies. g , Ratiometric sensor response curves for neutralizing antibody 47D11 (left) 44 , and commercially available anti-spike D001 (center) and D003 (right) 43 . Insets show sensor response at low target concentrations, which allowed calculation of the limit of detection, as indicated in the graphs. Experiments in b , d , e , g were performed in triplicate at sensor and calibrator concentrations as indicated in a , c , f , respectively. Reaction mixtures were prepared in buffer (PBS (pH 7.4), 0.1% (w/v) BSA) and incubated for 1 hr at room temperature before addition of NanoLuc substrate and recording of the emission spectra. The blue/green ratio was calculated by dividing bioluminescent emission at 458 nm by emission at 518 nm. The fold increase is indicated in each graph. Individual data points are represented as circles, while dashed lines connect mean values. Bars in histograms represent mean values.

    Article Snippet: Commercially available anti-SARS-COV-2 D001, D002, D003 and D004 were ordered from Sino Biological.

    Techniques: Binding Assay, Luciferase, Incubation

    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

    Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 1 1.0 and PiTou 2 3.0 furin prediction algorithm, generating a score with green numbers indicating predicted furin cleavage and red numbers indicating no predicted furin cleavage. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. Sequence numbers refer to position of amino acids within the spike protein. *For Bat-RmYN02, sequence number was determined from 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), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. Sequences corresponding to the S1/S2 and S2’ region of RmYN02 (EPI_ISL_412977) were obtained from GISAID.

    Journal: bioRxiv

    Article Title: Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage

    doi: 10.1101/2020.10.04.325522

    Figure Lengend Snippet: Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 1 1.0 and PiTou 2 3.0 furin prediction algorithm, generating a score with green numbers indicating predicted furin cleavage and red numbers indicating no predicted furin cleavage. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. Sequence numbers refer to position of amino acids within the spike protein. *For Bat-RmYN02, sequence number was determined from 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), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. Sequences corresponding to the S1/S2 and S2’ region of RmYN02 (EPI_ISL_412977) were obtained from GISAID.

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

    Techniques: Sequencing

    Predicted structural model of the SARS-CoV and SARS-CoV-2 S proteins. ( Inset ) Magnification of S1/S2 site with conserved R and S residues (red ribbon) and the unique four amino acid insertion P-R-R-A for SARS-CoV-2 (blue ribbon) are shown. The P’s denote the position of that amino acid from the S1/S2 cleavage site, with P1-P5 referring to amino acids before the cleavage site and P1’ referring to amino acids after the cleavage site.

    Journal: bioRxiv

    Article Title: Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage

    doi: 10.1101/2020.10.04.325522

    Figure Lengend Snippet: Predicted structural model of the SARS-CoV and SARS-CoV-2 S proteins. ( Inset ) Magnification of S1/S2 site with conserved R and S residues (red ribbon) and the unique four amino acid insertion P-R-R-A for SARS-CoV-2 (blue ribbon) are shown. The P’s denote the position of that amino acid from the S1/S2 cleavage site, with P1-P5 referring to amino acids before the cleavage site and P1’ referring to amino acids after the cleavage site.

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

    Techniques:

    Characterization of MLVpp system. ( A and 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-2pp and SARSpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region that cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLVp30. Image cropped from a singular Western blot and lanes are reshown in Figure 3C and 5C .

    Journal: bioRxiv

    Article Title: Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage

    doi: 10.1101/2020.10.04.325522

    Figure Lengend Snippet: Characterization of MLVpp system. ( A and 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-2pp and SARSpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region that cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLVp30. Image cropped from a singular Western blot and lanes are reshown in Figure 3C and 5C .

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

    Techniques: Infection, Luciferase, Activity Assay, Western Blot

    Analysis of the effect of Fr 1C on the SARS-CoV-2 proteins and genome. DMSO and Fr 1C were added to cell culture supernatants containing SARS-CoV-2 and were incubated at 25 °C for 48 h. n = 3 per group. ( A,B ) The images are the results of WB to detect SARS-CoV-2 ( A ) S2 subunit protein and ( B ) NP. ( C ) The image is the result of RT–PCR using NIID_2019-nCoV_N_F2 and R2 primers which amplify 158 bp region on SARS-CoV-2 gene. M: Marker.

    Journal: Viruses

    Article Title: Saxifraga spinulosa-Derived Components Rapidly Inactivate Multiple Viruses Including SARS-CoV-2

    doi: 10.3390/v12070699

    Figure Lengend Snippet: Analysis of the effect of Fr 1C on the SARS-CoV-2 proteins and genome. DMSO and Fr 1C were added to cell culture supernatants containing SARS-CoV-2 and were incubated at 25 °C for 48 h. n = 3 per group. ( A,B ) The images are the results of WB to detect SARS-CoV-2 ( A ) S2 subunit protein and ( B ) NP. ( C ) The image is the result of RT–PCR using NIID_2019-nCoV_N_F2 and R2 primers which amplify 158 bp region on SARS-CoV-2 gene. M: Marker.

    Article Snippet: SARS-CoV-2 S2 subunit protein was detected using rabbit anti-spike/S2 polyclonal antibody (Catalog No. 40590-T62, Sino Biological Inc.) and mouse anti-rabbit IgG peroxidase conjugate.

    Techniques: Cell Culture, Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Marker