sars cov s  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike RBD Gene ORF cDNA clone expression plasmid C His tag Codon Optimized COVID 19 Spike S RBD Research
    Description:
    Full length Clone DNA of SARS CoV 2 2019 nCoV Spike RBD
    Catalog Number:
    VG40592-CH
    Price:
    195.0
    Category:
    cDNA Clone
    Applications:
    Stable or Transient mammalian expression
    Size:
    1Unit
    Product Aliases:
    coronavirus spike cDNA ORF Clone 2019-nCoV, cov spike cDNA ORF Clone 2019-nCoV, ncov RBD cDNA ORF Clone 2019-nCoV, ncov s1 cDNA ORF Clone 2019-nCoV, ncov s2 cDNA ORF Clone 2019-nCoV, ncov spike cDNA ORF Clone 2019-nCoV, NCP-CoV RBD cDNA ORF Clone 2019-nCoV, NCP-CoV s1 cDNA ORF Clone 2019-nCoV, NCP-CoV s2 cDNA ORF Clone 2019-nCoV, NCP-CoV Spike cDNA ORF Clone 2019-nCoV, novel coronavirus RBD cDNA ORF Clone 2019-nCoV, novel coronavirus s1 cDNA ORF Clone 2019-nCoV, novel coronavirus s2 cDNA ORF Clone 2019-nCoV, novel coronavirus spike cDNA ORF Clone 2019-nCoV, RBD cDNA ORF Clone 2019-nCoV, S1 cDNA ORF Clone 2019-nCoV, S2 cDNA ORF Clone 2019-nCoV, Spike RBD cDNA ORF Clone 2019-nCoV
    Molecule Name:
    Spike
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    Structured Review

    Sino Biological sars cov s
    SARS CoV 2 2019 nCoV Spike RBD Gene ORF cDNA clone expression plasmid C His tag Codon Optimized COVID 19 Spike S RBD Research
    Full length Clone DNA of SARS CoV 2 2019 nCoV Spike RBD
    https://www.bioz.com/result/sars cov s/product/Sino Biological
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov s - by Bioz Stars, 2021-07
    99/100 stars

    Images

    1) Product Images from "Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage"

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

    Journal: bioRxiv

    doi: 10.1101/2020.10.04.325522

    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.
    Figure Legend 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.

    Techniques Used: Sequencing

    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 .
    Figure Legend 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 .

    Techniques Used: Infection, Luciferase, Activity Assay, Western Blot

    Related Articles

    Binding Assay:

    Article Title: Discovery of Aptamers Targeting the Receptor-Binding Domain of the SARS-CoV-2 Spike Glycoprotein
    Article Snippet: Flow cytometry results indicated that all the mutants showed weaker binding performance against RBD-Ni-beads than original aptamers ( C,G). .. This result indicated that MDS helps us to understand binding patterns at the molecular level of the two aptamers targeting RBD and suggests two aptamers may have partially identical binding sites, like ACE2 on SARS-CoV-2 RBD. .. Further study of the binding surface/sites between RBD and aptamers can help improve the relationship and enhance competitiveness with ACE2.

    Article Title: A Rapid and Efficient Screening System for Neutralizing Antibodies and Its Application for SARS-CoV-2
    Article Snippet: .. Moreover, we measured the binding affinity of top 20 NAbs to SARS-CoV-2 RBD via the surface plasmon resonance (SPR) ( ). .. These mAbs had equilibrium constant (KD ) values for the binding affinity ranging from 0.08 to 8 nM, except 81C3 (KD = 24 nM) ( ).

    other:

    Article Title: Discovery of Aptamers Targeting the Receptor-Binding Domain of the SARS-CoV-2 Spike Glycoprotein
    Article Snippet: Molecular Docking and Molecular Dynamic Simulations To better understand the interaction patterns between SARS-CoV-2 RBD and the two aptamers, we performed molecular docking and molecular dynamics simulations (MDS).

    Construct:

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct was derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and were used interchangeably. .. To construct a replication competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 spike protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene:1786) or an catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and was used interchangeably. .. To construct a replication-competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 S protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene; 1786) or a catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Sequencing:

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct was derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and were used interchangeably. .. To construct a replication competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 spike protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene:1786) or an catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and was used interchangeably. .. To construct a replication-competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 S protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene; 1786) or a catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Clone Assay:

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct was derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and were used interchangeably. .. To construct a replication competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 spike protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene:1786) or an catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and was used interchangeably. .. To construct a replication-competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 S protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene; 1786) or a catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Recombinant:

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct was derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and were used interchangeably. .. To construct a replication competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 spike protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene:1786) or an catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Article Title: An engineered stable mini-protein to plug SARS-Cov-2 Spikes
    Article Snippet: Microscale thermophoresis (MST) binding studies The thermophoretic measurements were performed using Monolith NT.115 device with red detection channel (NanoTemper Technologies, Munich, Germany). .. Recombinant Spikeplug was produced in our laboratory, while SARS-CoV-2 Spike RBD was purchased from Sino Biological Inc. For MST recording, the SARS-CoV-2 Spike RBD was labelled with the fluorescent dye NT647 using the protocol suggested from the NanoTemper. .. Thermophoretic experiments were conducted using Monolith NT.115 (NanoTemper Technologies, Munich, Germany).

    Article Title: Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses
    Article Snippet: A second construct derived from a codon-optimized plasmid (SinoBiological) behaved identically in our assays and was used interchangeably. .. To construct a replication-competent rVSV/SARS-CoV-2 chimeric virus clone, a codon-optimized cDNA sequence encoding the SARS-CoV-2 S protein (SinoBiological) but lacking the C-terminal 18 codons was inserted, using Gibson cloning, into a recombinant VSV background that contains GFP immediately upstream of the L (polymerase) following a strategy we previously described for the exchange of VSV-G with HIV-1 Env proteins ( ). .. An ACE2 lentivirus expression CS(ACE2)IB vector was constructed by inserting a cDNA encoding an unaltered ACE2 (Addgene; 1786) or a catalytically inactive ACE2 mutant (ACE2-H374N & H378N) into the lentivirus expression vector CSIB ( ).

    Produced:

    Article Title: An engineered stable mini-protein to plug SARS-Cov-2 Spikes
    Article Snippet: Microscale thermophoresis (MST) binding studies The thermophoretic measurements were performed using Monolith NT.115 device with red detection channel (NanoTemper Technologies, Munich, Germany). .. Recombinant Spikeplug was produced in our laboratory, while SARS-CoV-2 Spike RBD was purchased from Sino Biological Inc. For MST recording, the SARS-CoV-2 Spike RBD was labelled with the fluorescent dye NT647 using the protocol suggested from the NanoTemper. .. Thermophoretic experiments were conducted using Monolith NT.115 (NanoTemper Technologies, Munich, Germany).

    Plasmid Preparation:

    Article Title: Development of humanized tri-specific nanobodies with potent neutralization for SARS-CoV-2
    Article Snippet: .. Generation of deletion mutants of the SARS-CoV-2 S proteinThe codon-optimized version of the open reading frame cDNA for SARS-CoV-2 S protein was purchased from Sino Biological in the vector pCMV3-SP-N-Myc. .. The deletion mutants of the SARS-CoV-2 S were generated by the QuikChange II XL Site-Directed Mutagenesis Kit (Agilent Technologies, Cat.200522) according to manufacturer’s instructions.

    SPR Assay:

    Article Title: A Rapid and Efficient Screening System for Neutralizing Antibodies and Its Application for SARS-CoV-2
    Article Snippet: .. Moreover, we measured the binding affinity of top 20 NAbs to SARS-CoV-2 RBD via the surface plasmon resonance (SPR) ( ). .. These mAbs had equilibrium constant (KD ) values for the binding affinity ranging from 0.08 to 8 nM, except 81C3 (KD = 24 nM) ( ).

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    Sino Biological sars cov 2 rbd
    RU169 output clone diversity Using the <t>SARS-CoV-2</t> RBD as the target of library panning and FACS selection for screen RU169 produced a high number of unique clones, indicating high, unexplored, diversity in the output.
    Sars Cov 2 Rbd, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 rbd/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 rbd - by Bioz Stars, 2021-07
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    98
    Sino Biological sars cov 2 rbd protein
    Immunogenicity evaluation of a single mRNA-RBD vaccination. a – c Groups of BALB/c mice ( n = 6) were immunized with a single injection of mRNA-RBD at different doses or with a placebo via the i.m. route. Sera at 4 weeks post immunization were collected. <t>SARS-CoV-2</t> RBD-specific IgG ( a ) and neutralizing antibody titers in sera against pseudovirus ( b ) and live virus ( c ) infection were determined. d – h C57BL/6 mice ( n = 6) were inoculated with a single mRNA-RBD vaccination or a placebo. Serum samples were collected from mice at 4 weeks following vaccination. RBD-specific IgG titers and pseudovirus-neutralizing antibodies were measured as shown in d and e , respectively. f An ELISPOT assay was performed to evaluate the capacity of splenocytes to secrete IFNγ following re-stimulation with SARS-CoV-2 RBD peptide pools. g , h An ICS assay was conducted to quantify the proportions of IFNγ-secreting CD8 + ( g ) and CD4 + ( h ) T cells. mRNA-RBD-L indicates the low dose (2 μg). mRNA-RBD-H indicates the high dose (15 μg). HCS represents human convalescent sera. Data are means ± SEM (standard error of the mean). Comparisons were performed by Student’s t -test (unpaired, two tailed). Placebo animals = black circles; mRNA-RBD-L vaccinated animals = blue triangles; mRNA-RBD-H vaccinated animals = red squares; HCS = brown circles; dotted line = the limit of detection. Data are one representative result of two independent experiments. Source data are provided as a Source Data file.
    Sars Cov 2 Rbd Protein, 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 rbd protein/product/Sino Biological
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 rbd protein - by Bioz Stars, 2021-07
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    95
    Sino Biological sars cov 2
    Dual staining to detect <t>SARS-CoV-2</t> antigen and RNA in the same FFPE section. ( A–B ) Compared to uninfected control FFPE cell pellets ( A ), SARS-CoV-2 S (brown) and positive-sense RNA (red) were detected in the same section ( B ). Nuclei are stained blue (hematoxylin). Scale bar, 50 μm in ( A – B ).
    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
    https://www.bioz.com/result/sars cov 2/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 - by Bioz Stars, 2021-07
    95/100 stars
      Buy from Supplier

    Image Search Results


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

    Journal: bioRxiv

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

    doi: 10.1101/2020.07.27.224089

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

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

    Techniques: FACS, Selection, Produced, Clone Assay

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

    Journal: bioRxiv

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

    doi: 10.1101/2020.07.27.224089

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

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

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

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

    Journal: bioRxiv

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

    doi: 10.1101/2020.07.27.224089

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

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

    Techniques: Clone Assay, In Vitro

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

    Journal: bioRxiv

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

    doi: 10.1101/2020.07.27.224089

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

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

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

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

    Journal: bioRxiv

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

    doi: 10.1101/2020.07.27.224089

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

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

    Techniques: Binding Assay

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

    Journal: bioRxiv

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

    doi: 10.1101/2020.07.27.224089

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

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

    Techniques: Cytometry, Inhibition, Fluorescence, Labeling

    Immunogenicity evaluation of a single mRNA-RBD vaccination. a – c Groups of BALB/c mice ( n = 6) were immunized with a single injection of mRNA-RBD at different doses or with a placebo via the i.m. route. Sera at 4 weeks post immunization were collected. SARS-CoV-2 RBD-specific IgG ( a ) and neutralizing antibody titers in sera against pseudovirus ( b ) and live virus ( c ) infection were determined. d – h C57BL/6 mice ( n = 6) were inoculated with a single mRNA-RBD vaccination or a placebo. Serum samples were collected from mice at 4 weeks following vaccination. RBD-specific IgG titers and pseudovirus-neutralizing antibodies were measured as shown in d and e , respectively. f An ELISPOT assay was performed to evaluate the capacity of splenocytes to secrete IFNγ following re-stimulation with SARS-CoV-2 RBD peptide pools. g , h An ICS assay was conducted to quantify the proportions of IFNγ-secreting CD8 + ( g ) and CD4 + ( h ) T cells. mRNA-RBD-L indicates the low dose (2 μg). mRNA-RBD-H indicates the high dose (15 μg). HCS represents human convalescent sera. Data are means ± SEM (standard error of the mean). Comparisons were performed by Student’s t -test (unpaired, two tailed). Placebo animals = black circles; mRNA-RBD-L vaccinated animals = blue triangles; mRNA-RBD-H vaccinated animals = red squares; HCS = brown circles; dotted line = the limit of detection. Data are one representative result of two independent experiments. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: A single-dose mRNA vaccine provides a long-term protection for hACE2 transgenic mice from SARS-CoV-2

    doi: 10.1038/s41467-021-21037-2

    Figure Lengend Snippet: Immunogenicity evaluation of a single mRNA-RBD vaccination. a – c Groups of BALB/c mice ( n = 6) were immunized with a single injection of mRNA-RBD at different doses or with a placebo via the i.m. route. Sera at 4 weeks post immunization were collected. SARS-CoV-2 RBD-specific IgG ( a ) and neutralizing antibody titers in sera against pseudovirus ( b ) and live virus ( c ) infection were determined. d – h C57BL/6 mice ( n = 6) were inoculated with a single mRNA-RBD vaccination or a placebo. Serum samples were collected from mice at 4 weeks following vaccination. RBD-specific IgG titers and pseudovirus-neutralizing antibodies were measured as shown in d and e , respectively. f An ELISPOT assay was performed to evaluate the capacity of splenocytes to secrete IFNγ following re-stimulation with SARS-CoV-2 RBD peptide pools. g , h An ICS assay was conducted to quantify the proportions of IFNγ-secreting CD8 + ( g ) and CD4 + ( h ) T cells. mRNA-RBD-L indicates the low dose (2 μg). mRNA-RBD-H indicates the high dose (15 μg). HCS represents human convalescent sera. Data are means ± SEM (standard error of the mean). Comparisons were performed by Student’s t -test (unpaired, two tailed). Placebo animals = black circles; mRNA-RBD-L vaccinated animals = blue triangles; mRNA-RBD-H vaccinated animals = red squares; HCS = brown circles; dotted line = the limit of detection. Data are one representative result of two independent experiments. Source data are provided as a Source Data file.

    Article Snippet: Briefly, a monoclonal antibody specific for SARS-CoV-2 RBD protein was pre-coated onto plate wells.

    Techniques: Mouse Assay, Injection, Infection, Enzyme-linked Immunospot, Two Tailed Test

    Duration and long-term protection of humoral response induced by mRNA-RBD. a Passive immunization and challenge schedule. The blue and red arrow indicates the time of vaccination and sera transfer, respectively. b , c Groups of BALB/c mice ( n = 10) received 15 μg of mRNA-RBD or a placebo. Half of the mice per group were euthanized at 8 weeks (short term) post vaccination, and massive sera were collected for further passive immunization. The other mice of the group were bled as desired and eventually euthanized at 26 weeks (long term) post vaccination to collect massive sera for further passive immunization. All serum samples were detected for IgG ( b ) and neutralizing antibodies ( c ) titers. d–e hACE2 transgenic mice ( n = 5) were administered 350 μl per mouse of pooled short- and long-term immune sera and one day later were challenged with 1 × 10 5 FFU of SARS-CoV-2 via the i.n. route. d The hACE2 mice weight change was recorded after challenge. e Virus titers in lung. mRNA-RBD-H indicates the high-dose vaccine (15 μg). Data are means ± SEM (standard error of the mean). Comparisons were performed by Student’s t -test (unpaired, two tailed). Placebo animals = black circles; animals for long-term study = blue triangles; animals for short-term study = red squares; dotted line = the limit of detection. Data are one representative result of two independent experiments. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: A single-dose mRNA vaccine provides a long-term protection for hACE2 transgenic mice from SARS-CoV-2

    doi: 10.1038/s41467-021-21037-2

    Figure Lengend Snippet: Duration and long-term protection of humoral response induced by mRNA-RBD. a Passive immunization and challenge schedule. The blue and red arrow indicates the time of vaccination and sera transfer, respectively. b , c Groups of BALB/c mice ( n = 10) received 15 μg of mRNA-RBD or a placebo. Half of the mice per group were euthanized at 8 weeks (short term) post vaccination, and massive sera were collected for further passive immunization. The other mice of the group were bled as desired and eventually euthanized at 26 weeks (long term) post vaccination to collect massive sera for further passive immunization. All serum samples were detected for IgG ( b ) and neutralizing antibodies ( c ) titers. d–e hACE2 transgenic mice ( n = 5) were administered 350 μl per mouse of pooled short- and long-term immune sera and one day later were challenged with 1 × 10 5 FFU of SARS-CoV-2 via the i.n. route. d The hACE2 mice weight change was recorded after challenge. e Virus titers in lung. mRNA-RBD-H indicates the high-dose vaccine (15 μg). Data are means ± SEM (standard error of the mean). Comparisons were performed by Student’s t -test (unpaired, two tailed). Placebo animals = black circles; animals for long-term study = blue triangles; animals for short-term study = red squares; dotted line = the limit of detection. Data are one representative result of two independent experiments. Source data are provided as a Source Data file.

    Article Snippet: Briefly, a monoclonal antibody specific for SARS-CoV-2 RBD protein was pre-coated onto plate wells.

    Techniques: Mouse Assay, Transgenic Assay, Two Tailed Test

    Protection efficacy of mRNA-RBD in hACE2 transgenic mice against SARS-CoV-2. a-d Groups of hACE2 transgenic mice ( n = 6) received one (prime group) or two (boost group) doses of mRNA-RBD-H or placebo via the i.m. route. Four weeks post initial vaccination, mice were challenged with 1 × 10 5 FFU of SARS-CoV-2 virus. a Mice immunization and challenge schedule. The blue arrows indicate the time of vaccination. b , c Sera collected at 4 weeks post initial vaccination were examined for IgG ( b ) and neutralizing antibody ( c ) titers. d Mice weight change after challenge. e Virus titers in lungs of challenged mice ( n = 4). f Representative histopathology (H E) of lungs in SARS-CoV-2-infected hACE2 mice (5 dpi). Infiltration of lymphocytes within alveolar spaces is indicated by yellow arrows. Scale bar, 100 μm. g Representative immunohistochemistry (IHC) of lung tissues with SARS-CoV-2 N-specific monoclonal antibodies. Virus is indicated by yellow arrows. Scale bar, 100 μm. mRNA-RBD-H indicates the high-dose vaccine (15 μg). Data are means ± SEM (standard error of the mean). Comparisons were performed by Student’s t -test (unpaired, two tailed). Placebo animals = black circles; one injection-animals = blue triangles; two injections-vaccinated animals = red squares; dotted line = the limit of detection. Data are one representative result of two independent experiments. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: A single-dose mRNA vaccine provides a long-term protection for hACE2 transgenic mice from SARS-CoV-2

    doi: 10.1038/s41467-021-21037-2

    Figure Lengend Snippet: Protection efficacy of mRNA-RBD in hACE2 transgenic mice against SARS-CoV-2. a-d Groups of hACE2 transgenic mice ( n = 6) received one (prime group) or two (boost group) doses of mRNA-RBD-H or placebo via the i.m. route. Four weeks post initial vaccination, mice were challenged with 1 × 10 5 FFU of SARS-CoV-2 virus. a Mice immunization and challenge schedule. The blue arrows indicate the time of vaccination. b , c Sera collected at 4 weeks post initial vaccination were examined for IgG ( b ) and neutralizing antibody ( c ) titers. d Mice weight change after challenge. e Virus titers in lungs of challenged mice ( n = 4). f Representative histopathology (H E) of lungs in SARS-CoV-2-infected hACE2 mice (5 dpi). Infiltration of lymphocytes within alveolar spaces is indicated by yellow arrows. Scale bar, 100 μm. g Representative immunohistochemistry (IHC) of lung tissues with SARS-CoV-2 N-specific monoclonal antibodies. Virus is indicated by yellow arrows. Scale bar, 100 μm. mRNA-RBD-H indicates the high-dose vaccine (15 μg). Data are means ± SEM (standard error of the mean). Comparisons were performed by Student’s t -test (unpaired, two tailed). Placebo animals = black circles; one injection-animals = blue triangles; two injections-vaccinated animals = red squares; dotted line = the limit of detection. Data are one representative result of two independent experiments. Source data are provided as a Source Data file.

    Article Snippet: Briefly, a monoclonal antibody specific for SARS-CoV-2 RBD protein was pre-coated onto plate wells.

    Techniques: Transgenic Assay, Mouse Assay, Histopathology, Infection, Immunohistochemistry, Two Tailed Test, Injection

    Construction and characterization of mRNA-RBD vaccine. a Schematic of the mRNA-RBD vaccine design. The SARS-CoV-2 mRNA encodes the signal peptide (SP), receptor-binding domain (RBD) from SARS-CoV-2 strain Wuhan/IVDC-HB-01/2019. b mRNA-RBD was transfected into HEK293T cells. RBD expression in the cell lysate and supernatant was analyzed by western blotting. c Particle size of LNPs by dynamic light scattering. d A representative cryo-electron microscopy image of a LNPs solution following mRNA encapsulation. Scale bar, 100 nm. e Zeta potential for LNPs at pH 4.0 and 7.4. For b and d , two independent experiments were carried out with similar results. For c and e , one representative result from three independent experiments is shown. Source data are provided as a Source Data file.

    Journal: Nature Communications

    Article Title: A single-dose mRNA vaccine provides a long-term protection for hACE2 transgenic mice from SARS-CoV-2

    doi: 10.1038/s41467-021-21037-2

    Figure Lengend Snippet: Construction and characterization of mRNA-RBD vaccine. a Schematic of the mRNA-RBD vaccine design. The SARS-CoV-2 mRNA encodes the signal peptide (SP), receptor-binding domain (RBD) from SARS-CoV-2 strain Wuhan/IVDC-HB-01/2019. b mRNA-RBD was transfected into HEK293T cells. RBD expression in the cell lysate and supernatant was analyzed by western blotting. c Particle size of LNPs by dynamic light scattering. d A representative cryo-electron microscopy image of a LNPs solution following mRNA encapsulation. Scale bar, 100 nm. e Zeta potential for LNPs at pH 4.0 and 7.4. For b and d , two independent experiments were carried out with similar results. For c and e , one representative result from three independent experiments is shown. Source data are provided as a Source Data file.

    Article Snippet: Briefly, a monoclonal antibody specific for SARS-CoV-2 RBD protein was pre-coated onto plate wells.

    Techniques: Binding Assay, Transfection, Expressing, Western Blot, Electron Microscopy

    Structural analysis of P4A1 Fab and SARS-CoV-2 RBD complex. a The overall P4A1-Fab-RBD complex structure superimposed with the hACE2-RBD complex. The P4A1 heavy chain (colored slate blue), light chain (colored salmon red), and hACE (colored pale green) are displayed in cartoon representation. The SARS-CoV-2 RBD is colored in gray and displayed in surface representation. b The epitope of P4A1 shown in surface representation. The CDR loops of heavy chain (HCDR) and light chain (LCDR) are colored in purple and magenta, respectively. The epitopes from the heavy chain and light chain are colored in slate blue and salmon red, respectively. The only residue K417, which contacts with both heavy chain and light chain, is colored in pink. The light-chain frame region 3 (LFR3) is colored in orange. The identical residues on RBD shared in P4A1 and hACE2 binding are labeled in red. The residues are numbered according to SARS-CoV-2 RBD. c The detailed interactions between SARS-CoV-2 RBD with HCDR, LCDR, and LFR3. The residues are shown in sticks with identical colors to ( b ).

    Journal: Nature Communications

    Article Title: A SARS-CoV-2 neutralizing antibody with extensive Spike binding coverage and modified for optimal therapeutic outcomes

    doi: 10.1038/s41467-021-22926-2

    Figure Lengend Snippet: Structural analysis of P4A1 Fab and SARS-CoV-2 RBD complex. a The overall P4A1-Fab-RBD complex structure superimposed with the hACE2-RBD complex. The P4A1 heavy chain (colored slate blue), light chain (colored salmon red), and hACE (colored pale green) are displayed in cartoon representation. The SARS-CoV-2 RBD is colored in gray and displayed in surface representation. b The epitope of P4A1 shown in surface representation. The CDR loops of heavy chain (HCDR) and light chain (LCDR) are colored in purple and magenta, respectively. The epitopes from the heavy chain and light chain are colored in slate blue and salmon red, respectively. The only residue K417, which contacts with both heavy chain and light chain, is colored in pink. The light-chain frame region 3 (LFR3) is colored in orange. The identical residues on RBD shared in P4A1 and hACE2 binding are labeled in red. The residues are numbered according to SARS-CoV-2 RBD. c The detailed interactions between SARS-CoV-2 RBD with HCDR, LCDR, and LFR3. The residues are shown in sticks with identical colors to ( b ).

    Article Snippet: Serially diluted SARS-CoV-2 RBD (WT, AcroBiosystems, Catalog #SPD-C52H3), RBD (V367F, AcroBiosystems, Catalog #SPD-S52H4), RBD (N354D/D364Y, AcroBiosystems, Catalog #SPD-S52H3), RBD (R408I, AcroBiosystems, Catalog #SPD-S52H8), RBD (W436R, AcroBiosystems, Catalog #SPD-S52H7), RBD (N439K, Sino Biological, Catalog #40592-V08H14), RBD (Y453F, Sino Biological, Catalog #40592-V08H80), RBD (S477N, Sino Biological, Catalog #40592-V08H46), RBD (F490S, Sino Biological, Catalog #40592-V08H41), RBD (S494P, Sino Biological, Catalog #40592-V08H18), RBD (N501Y, Sino Biological, Catalog #40592-V08H82), or SARS-CoV-2 spike S1 domain (D614G, Sino Biological, Catalog #40591-V08H3) were injected through flow cells for 60 or 180 s of association followed by a 150 or 800 s dissociation phase at a flow rate of 30 μL min−1 .

    Techniques: Binding Assay, Labeling

    Therapeutic efficacy of in the rhesus macaque model of SARS-CoV-2 infection. a Experimental design for therapeutic testing of P4A1–2A in the rhesus macaque ( n = 3/group). b Viral load in oropharyngeal swabs tested by RT-qPCR was monitored for 7 days. c Viral load in the respiratory tissues (including trachea, left and right bronchus, and all six lung lobes) collected at necropsy on 7 days post infection (d.p.i., n = 1/group) was tested by RT-qPCR. d Representative images of histopathology in lung tissue from isotype control or P4A1–2A 50 mg/kg treated animals (collected at 7 d.p.i., n = 1/group).

    Journal: Nature Communications

    Article Title: A SARS-CoV-2 neutralizing antibody with extensive Spike binding coverage and modified for optimal therapeutic outcomes

    doi: 10.1038/s41467-021-22926-2

    Figure Lengend Snippet: Therapeutic efficacy of in the rhesus macaque model of SARS-CoV-2 infection. a Experimental design for therapeutic testing of P4A1–2A in the rhesus macaque ( n = 3/group). b Viral load in oropharyngeal swabs tested by RT-qPCR was monitored for 7 days. c Viral load in the respiratory tissues (including trachea, left and right bronchus, and all six lung lobes) collected at necropsy on 7 days post infection (d.p.i., n = 1/group) was tested by RT-qPCR. d Representative images of histopathology in lung tissue from isotype control or P4A1–2A 50 mg/kg treated animals (collected at 7 d.p.i., n = 1/group).

    Article Snippet: Serially diluted SARS-CoV-2 RBD (WT, AcroBiosystems, Catalog #SPD-C52H3), RBD (V367F, AcroBiosystems, Catalog #SPD-S52H4), RBD (N354D/D364Y, AcroBiosystems, Catalog #SPD-S52H3), RBD (R408I, AcroBiosystems, Catalog #SPD-S52H8), RBD (W436R, AcroBiosystems, Catalog #SPD-S52H7), RBD (N439K, Sino Biological, Catalog #40592-V08H14), RBD (Y453F, Sino Biological, Catalog #40592-V08H80), RBD (S477N, Sino Biological, Catalog #40592-V08H46), RBD (F490S, Sino Biological, Catalog #40592-V08H41), RBD (S494P, Sino Biological, Catalog #40592-V08H18), RBD (N501Y, Sino Biological, Catalog #40592-V08H82), or SARS-CoV-2 spike S1 domain (D614G, Sino Biological, Catalog #40591-V08H3) were injected through flow cells for 60 or 180 s of association followed by a 150 or 800 s dissociation phase at a flow rate of 30 μL min−1 .

    Techniques: Infection, Quantitative RT-PCR, Histopathology

    The activities of IgG4 antibody P4A1–2A to different SARS-CoV-2 S protein mutants, FcRs, and C1q. a Binding of antibody P4A1 to SARS-CoV-2 S protein N354D/D364Y, R408I, W436R, V367F, or D614G mutants determined by surface plasmon resonance (SPR). b Pseudovirus neutralization assay in hACE2-overexpressing HEK293 cells. Experiment performed in triplicates with symbols represent each of the triplicates. c The binding affinity of P4A1 and P4A1–2A for different human FcRs and complement C1q.

    Journal: Nature Communications

    Article Title: A SARS-CoV-2 neutralizing antibody with extensive Spike binding coverage and modified for optimal therapeutic outcomes

    doi: 10.1038/s41467-021-22926-2

    Figure Lengend Snippet: The activities of IgG4 antibody P4A1–2A to different SARS-CoV-2 S protein mutants, FcRs, and C1q. a Binding of antibody P4A1 to SARS-CoV-2 S protein N354D/D364Y, R408I, W436R, V367F, or D614G mutants determined by surface plasmon resonance (SPR). b Pseudovirus neutralization assay in hACE2-overexpressing HEK293 cells. Experiment performed in triplicates with symbols represent each of the triplicates. c The binding affinity of P4A1 and P4A1–2A for different human FcRs and complement C1q.

    Article Snippet: Serially diluted SARS-CoV-2 RBD (WT, AcroBiosystems, Catalog #SPD-C52H3), RBD (V367F, AcroBiosystems, Catalog #SPD-S52H4), RBD (N354D/D364Y, AcroBiosystems, Catalog #SPD-S52H3), RBD (R408I, AcroBiosystems, Catalog #SPD-S52H8), RBD (W436R, AcroBiosystems, Catalog #SPD-S52H7), RBD (N439K, Sino Biological, Catalog #40592-V08H14), RBD (Y453F, Sino Biological, Catalog #40592-V08H80), RBD (S477N, Sino Biological, Catalog #40592-V08H46), RBD (F490S, Sino Biological, Catalog #40592-V08H41), RBD (S494P, Sino Biological, Catalog #40592-V08H18), RBD (N501Y, Sino Biological, Catalog #40592-V08H82), or SARS-CoV-2 spike S1 domain (D614G, Sino Biological, Catalog #40591-V08H3) were injected through flow cells for 60 or 180 s of association followed by a 150 or 800 s dissociation phase at a flow rate of 30 μL min−1 .

    Techniques: Binding Assay, SPR Assay, Neutralization

    Characterization of neutralizing antibodies from convalescent patients. a Characterization of SARS-CoV-2 S protein-specific antibodies. Upper panels: binding of antibodies to the full-length S protein, S1 protein, and S2 protein was evaluated by ELISA (in duplicates with symbols show each of the replicates). Lower left panel: blockage of the binding of SARS-CoV-2 Spike S1 protein to Vero E6 cells by antibodies evaluated by flow cytometry (data in singleton). Lower middle panel: pseudovirus neutralization assay in Huh-7 cells (data in singleton). Lower right panel: in triplicates with symbols show each of the triplicates and SARS-CoV-2 live virus neutralization assay. All experiments were repeated at least two more times (except S2 binding that was repeated one more time) with similar results. b Images of Vero E6 cell-infected SARS-CoV-2 treated with antibodies of different concentrations. Green (stained with SARS-CoV-2 nucleocapsid protein (NP) antibody) indicates viral infected cells and blue (Hoechst 33258) represents cell nuclei. Experiment was performed in triplicates and repeated two more times with similar results.

    Journal: Nature Communications

    Article Title: A SARS-CoV-2 neutralizing antibody with extensive Spike binding coverage and modified for optimal therapeutic outcomes

    doi: 10.1038/s41467-021-22926-2

    Figure Lengend Snippet: Characterization of neutralizing antibodies from convalescent patients. a Characterization of SARS-CoV-2 S protein-specific antibodies. Upper panels: binding of antibodies to the full-length S protein, S1 protein, and S2 protein was evaluated by ELISA (in duplicates with symbols show each of the replicates). Lower left panel: blockage of the binding of SARS-CoV-2 Spike S1 protein to Vero E6 cells by antibodies evaluated by flow cytometry (data in singleton). Lower middle panel: pseudovirus neutralization assay in Huh-7 cells (data in singleton). Lower right panel: in triplicates with symbols show each of the triplicates and SARS-CoV-2 live virus neutralization assay. All experiments were repeated at least two more times (except S2 binding that was repeated one more time) with similar results. b Images of Vero E6 cell-infected SARS-CoV-2 treated with antibodies of different concentrations. Green (stained with SARS-CoV-2 nucleocapsid protein (NP) antibody) indicates viral infected cells and blue (Hoechst 33258) represents cell nuclei. Experiment was performed in triplicates and repeated two more times with similar results.

    Article Snippet: Serially diluted SARS-CoV-2 RBD (WT, AcroBiosystems, Catalog #SPD-C52H3), RBD (V367F, AcroBiosystems, Catalog #SPD-S52H4), RBD (N354D/D364Y, AcroBiosystems, Catalog #SPD-S52H3), RBD (R408I, AcroBiosystems, Catalog #SPD-S52H8), RBD (W436R, AcroBiosystems, Catalog #SPD-S52H7), RBD (N439K, Sino Biological, Catalog #40592-V08H14), RBD (Y453F, Sino Biological, Catalog #40592-V08H80), RBD (S477N, Sino Biological, Catalog #40592-V08H46), RBD (F490S, Sino Biological, Catalog #40592-V08H41), RBD (S494P, Sino Biological, Catalog #40592-V08H18), RBD (N501Y, Sino Biological, Catalog #40592-V08H82), or SARS-CoV-2 spike S1 domain (D614G, Sino Biological, Catalog #40591-V08H3) were injected through flow cells for 60 or 180 s of association followed by a 150 or 800 s dissociation phase at a flow rate of 30 μL min−1 .

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Neutralization, Infection, Staining

    Dual staining to detect SARS-CoV-2 antigen and RNA in the same FFPE section. ( A–B ) Compared to uninfected control FFPE cell pellets ( A ), SARS-CoV-2 S (brown) and positive-sense RNA (red) were detected in the same section ( B ). Nuclei are stained blue (hematoxylin). Scale bar, 50 μm in ( A – B ).

    Journal: bioRxiv

    Article Title: Molecular Detection of SARS-CoV-2 in Formalin Fixed Paraffin Embedded Specimens

    doi: 10.1101/2020.04.21.042911

    Figure Lengend Snippet: Dual staining to detect SARS-CoV-2 antigen and RNA in the same FFPE section. ( A–B ) Compared to uninfected control FFPE cell pellets ( A ), SARS-CoV-2 S (brown) and positive-sense RNA (red) were detected in the same section ( B ). Nuclei are stained blue (hematoxylin). Scale bar, 50 μm in ( A – B ).

    Article Snippet: Identification of antibodies suitable for detection of SARS-CoV-2 by IHC and IFA in FFPE specimensTo identify antibodies that can be used to detect SARS-CoV-2 in human and animal tissues, we searched for commercially available SARS-CoV antibodies that recognize epitopes that are likely conserved in SARS-CoV-2.

    Techniques: Staining, Formalin-fixed Paraffin-Embedded

    Detection of SARS-CoV-2 replication in FFPE cells using multiplex fluorescence ISH. ( A–B ) Compared to uninfected control ( A ), SARS-CoV-2 negative-sense RNA (green), a replicative intermediate that indicates viral replication, can be detected in infected FFPE cell pellets in addition to positive-sense (red) RNA ( B ). Nuclei are stained blue (DAPI). Scale bar, 20 μm in ( A – B ).

    Journal: bioRxiv

    Article Title: Molecular Detection of SARS-CoV-2 in Formalin Fixed Paraffin Embedded Specimens

    doi: 10.1101/2020.04.21.042911

    Figure Lengend Snippet: Detection of SARS-CoV-2 replication in FFPE cells using multiplex fluorescence ISH. ( A–B ) Compared to uninfected control ( A ), SARS-CoV-2 negative-sense RNA (green), a replicative intermediate that indicates viral replication, can be detected in infected FFPE cell pellets in addition to positive-sense (red) RNA ( B ). Nuclei are stained blue (DAPI). Scale bar, 20 μm in ( A – B ).

    Article Snippet: Identification of antibodies suitable for detection of SARS-CoV-2 by IHC and IFA in FFPE specimensTo identify antibodies that can be used to detect SARS-CoV-2 in human and animal tissues, we searched for commercially available SARS-CoV antibodies that recognize epitopes that are likely conserved in SARS-CoV-2.

    Techniques: Formalin-fixed Paraffin-Embedded, Multiplex Assay, Fluorescence, In Situ Hybridization, Infection, Staining

    Detection of SARS-CoV-2 antigens by IHC and IFA in FFPE cell pellets. ( A–B ) In comparison to uninfected control FFPE cell pellets ( A and C ), SARS-CoV-2 S (brown, B ) and SARS-CoV-2 NP (brown, D ) can be detected in FFPE SARS-CoV-2-infected cell pellets. Nuclei are stained blue (hematoxylin). ( E ) Immunofluorescence staining to detect SARS-CoV-2 S (green) and NP (red) in FFPE SARS-CoV-2-infected cell pellets. Inset of ( E ) is uninfected control FFPE cell pellets. Nuclei are stained blue (DAPI). Scale bar, 50 μm in ( A – D ), 20 μm in inset of ( E) , and 10 μm in ( E) .

    Journal: bioRxiv

    Article Title: Molecular Detection of SARS-CoV-2 in Formalin Fixed Paraffin Embedded Specimens

    doi: 10.1101/2020.04.21.042911

    Figure Lengend Snippet: Detection of SARS-CoV-2 antigens by IHC and IFA in FFPE cell pellets. ( A–B ) In comparison to uninfected control FFPE cell pellets ( A and C ), SARS-CoV-2 S (brown, B ) and SARS-CoV-2 NP (brown, D ) can be detected in FFPE SARS-CoV-2-infected cell pellets. Nuclei are stained blue (hematoxylin). ( E ) Immunofluorescence staining to detect SARS-CoV-2 S (green) and NP (red) in FFPE SARS-CoV-2-infected cell pellets. Inset of ( E ) is uninfected control FFPE cell pellets. Nuclei are stained blue (DAPI). Scale bar, 50 μm in ( A – D ), 20 μm in inset of ( E) , and 10 μm in ( E) .

    Article Snippet: Identification of antibodies suitable for detection of SARS-CoV-2 by IHC and IFA in FFPE specimensTo identify antibodies that can be used to detect SARS-CoV-2 in human and animal tissues, we searched for commercially available SARS-CoV antibodies that recognize epitopes that are likely conserved in SARS-CoV-2.

    Techniques: Immunohistochemistry, Immunofluorescence, Formalin-fixed Paraffin-Embedded, Infection, Staining

    Detection of SARS-CoV-2 RNA by ISH in FFPE cell pellets. ( A–B ) SARS-CoV-2 positive-sense RNA can be detected by ISH using positive-sense RNA probe 1 in infected FFPE cell pellets ( B ), but not in uninfected control FFPE cell pellets ( A ). ( C–D ) SARS-CoV-2 positive-sense RNA can be detected by ISH using positive-sense RNA probe 2 in infected FFPE cell pellets ( D ), but not in uninfected control FFPE cell pellets ( C ). ( E–F ) SARS-CoV-2 negative-sense RNA can be detected by ISH using negative-sense RNA probe 1in infected FFPE cell pellets ( E ), but not in uninfected control FFPE cell pellets ( F ). Nuclei are stained blue (hematoxylin). Scale bar, 50 μm in ( A – F ).

    Journal: bioRxiv

    Article Title: Molecular Detection of SARS-CoV-2 in Formalin Fixed Paraffin Embedded Specimens

    doi: 10.1101/2020.04.21.042911

    Figure Lengend Snippet: Detection of SARS-CoV-2 RNA by ISH in FFPE cell pellets. ( A–B ) SARS-CoV-2 positive-sense RNA can be detected by ISH using positive-sense RNA probe 1 in infected FFPE cell pellets ( B ), but not in uninfected control FFPE cell pellets ( A ). ( C–D ) SARS-CoV-2 positive-sense RNA can be detected by ISH using positive-sense RNA probe 2 in infected FFPE cell pellets ( D ), but not in uninfected control FFPE cell pellets ( C ). ( E–F ) SARS-CoV-2 negative-sense RNA can be detected by ISH using negative-sense RNA probe 1in infected FFPE cell pellets ( E ), but not in uninfected control FFPE cell pellets ( F ). Nuclei are stained blue (hematoxylin). Scale bar, 50 μm in ( A – F ).

    Article Snippet: Identification of antibodies suitable for detection of SARS-CoV-2 by IHC and IFA in FFPE specimensTo identify antibodies that can be used to detect SARS-CoV-2 in human and animal tissues, we searched for commercially available SARS-CoV antibodies that recognize epitopes that are likely conserved in SARS-CoV-2.

    Techniques: In Situ Hybridization, Formalin-fixed Paraffin-Embedded, Infection, Staining