sars cov 2 rbd protein  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike RBD V341I His Recombinant Protein
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike RBD V341I His Recombinant Protein YP 009724390 1 Arg319 Phe541 V341I was expressed with a polyhistidine tag at the C terminus
    Catalog Number:
    40592-V08H11
    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:
    HEK293 Cells
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    Structured Review

    Sino Biological sars cov 2 rbd protein
    HumVHH_S56A/LALA-Fc/Gen2 neutralizes <t>SARS-CoV-2</t> variants of concern. a . Surface view of SARS-CoV-2 RBD (grey) with VHH72 (green cartoon, bottom) and the N-terminal helixes of ACE2 (blue cartoon, top). The RBD-residues K417, N439, E484 and N501 (orange) are indicated. b. Binding of humVHH_S56A/LALA-Fc/Gen2 (left), CB6 (middle) and palivizumab (right) to SARS-CoV-1 spike with the RBD replaced by WT, N439K, K417N, E484K, N501Y or (K417N + E484K + N501Y) RBD of SARS-CoV-2, expressed on the surface of 293T cells. Data points represent the ratio of the mean fluorescence intensity (MFI) of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells, as determined by flow cytometry. c . SARS-CoV-2 plaque reduction neutralization assay with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs using BetaCov/Belgium/GHB-03021/2020, B1.1.7, or B.1.351 variant viruses.
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike RBD V341I His Recombinant Protein YP 009724390 1 Arg319 Phe541 V341I was expressed with a polyhistidine tag at the C terminus
    https://www.bioz.com/result/sars cov 2 rbd protein/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 rbd protein - by Bioz Stars, 2021-05
    96/100 stars

    Images

    1) Product Images from "Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters"

    Article Title: Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters

    Journal: bioRxiv

    doi: 10.1101/2021.03.08.433449

    HumVHH_S56A/LALA-Fc/Gen2 neutralizes SARS-CoV-2 variants of concern. a . Surface view of SARS-CoV-2 RBD (grey) with VHH72 (green cartoon, bottom) and the N-terminal helixes of ACE2 (blue cartoon, top). The RBD-residues K417, N439, E484 and N501 (orange) are indicated. b. Binding of humVHH_S56A/LALA-Fc/Gen2 (left), CB6 (middle) and palivizumab (right) to SARS-CoV-1 spike with the RBD replaced by WT, N439K, K417N, E484K, N501Y or (K417N + E484K + N501Y) RBD of SARS-CoV-2, expressed on the surface of 293T cells. Data points represent the ratio of the mean fluorescence intensity (MFI) of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells, as determined by flow cytometry. c . SARS-CoV-2 plaque reduction neutralization assay with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs using BetaCov/Belgium/GHB-03021/2020, B1.1.7, or B.1.351 variant viruses.
    Figure Legend Snippet: HumVHH_S56A/LALA-Fc/Gen2 neutralizes SARS-CoV-2 variants of concern. a . Surface view of SARS-CoV-2 RBD (grey) with VHH72 (green cartoon, bottom) and the N-terminal helixes of ACE2 (blue cartoon, top). The RBD-residues K417, N439, E484 and N501 (orange) are indicated. b. Binding of humVHH_S56A/LALA-Fc/Gen2 (left), CB6 (middle) and palivizumab (right) to SARS-CoV-1 spike with the RBD replaced by WT, N439K, K417N, E484K, N501Y or (K417N + E484K + N501Y) RBD of SARS-CoV-2, expressed on the surface of 293T cells. Data points represent the ratio of the mean fluorescence intensity (MFI) of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells, as determined by flow cytometry. c . SARS-CoV-2 plaque reduction neutralization assay with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs using BetaCov/Belgium/GHB-03021/2020, B1.1.7, or B.1.351 variant viruses.

    Techniques Used: Binding Assay, Fluorescence, Transfection, Flow Cytometry, Neutralization, Construct, Variant Assay

    Therapeutic administration of VHH72-Fc constructs restricts SARS-CoV-2 virus replication in Syrian hamsters. a-c. Hamsters were challenged with 1×10 4 PFU of BetaCoV/Munich/BavPat1/2020 and 4 hours later injected intraperitoneally with 20, 7 or 2 mg/kg of bivalent humVHH_S56A/LALAPG-Fc/Gen2 or tetravalent (humVHH_S56A) 2 /LALAPG-Fc/Gen2. The negative control group was treated with 20 mg/kg of palivizumab, injected 4 hours after the challenge infection; hamsters in a prophylactic control group received 20 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day before the challenge. ( a ) Lung virus loads, ( b ) lung viral RNA copies, and ( c ) gross lung pathology determined on day 4 after infection. d,e . Hamsters received an intraperitoneal injection of 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day prior to challenge or were treated by intraperitoneal injection of 1 or 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 or (humVHH_S56A) 2 /LALAPG-Fc/Gen2 19h after infection with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Seven mg/kg of palivizumab was used as a negative control treatment. ( d ) Virus load and ( e ) viral RNA levels in the lungs on day 4 after challenge. f,g . Hamsters were treated with 4mg/kg of palivizumab, humVHH_S56A/LALA-Fc/Gen2 or humVHH/LALA-Fc/Gen2 injected intraperitoneally 24h after challenge with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Viral RNA and infectious virus were determined in lung tissue on day 4 after infection. Data were analyzed with the Mann-Whitney U-test using GraphPad Prism software. *, p
    Figure Legend Snippet: Therapeutic administration of VHH72-Fc constructs restricts SARS-CoV-2 virus replication in Syrian hamsters. a-c. Hamsters were challenged with 1×10 4 PFU of BetaCoV/Munich/BavPat1/2020 and 4 hours later injected intraperitoneally with 20, 7 or 2 mg/kg of bivalent humVHH_S56A/LALAPG-Fc/Gen2 or tetravalent (humVHH_S56A) 2 /LALAPG-Fc/Gen2. The negative control group was treated with 20 mg/kg of palivizumab, injected 4 hours after the challenge infection; hamsters in a prophylactic control group received 20 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day before the challenge. ( a ) Lung virus loads, ( b ) lung viral RNA copies, and ( c ) gross lung pathology determined on day 4 after infection. d,e . Hamsters received an intraperitoneal injection of 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day prior to challenge or were treated by intraperitoneal injection of 1 or 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 or (humVHH_S56A) 2 /LALAPG-Fc/Gen2 19h after infection with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Seven mg/kg of palivizumab was used as a negative control treatment. ( d ) Virus load and ( e ) viral RNA levels in the lungs on day 4 after challenge. f,g . Hamsters were treated with 4mg/kg of palivizumab, humVHH_S56A/LALA-Fc/Gen2 or humVHH/LALA-Fc/Gen2 injected intraperitoneally 24h after challenge with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Viral RNA and infectious virus were determined in lung tissue on day 4 after infection. Data were analyzed with the Mann-Whitney U-test using GraphPad Prism software. *, p

    Techniques Used: Construct, Injection, Negative Control, Infection, MANN-WHITNEY, Software

    Enhanced affinity and neutralizing activity of computationally predicted VHH72 variant. a-c. Prototype WT-VHH/12GS-WT-Fc protects K18-hACE2 transgenic mice against SARS-CoV-2 challenge. Mice were injected intraperitoneally with 5 mg/kg of either WT-VHH/12GS-WT-Fc (n = 17) or control M2e-VHH-Fc (directed against the matrix protein 2 ectodomain of influenza A; n = 10). Seven hours later, the mice were challenged with 8 x 10 3 PFU of a clinical SARS-CoV-2 isolate. ( a ) Body weight (symbols represent means ± SD, p
    Figure Legend Snippet: Enhanced affinity and neutralizing activity of computationally predicted VHH72 variant. a-c. Prototype WT-VHH/12GS-WT-Fc protects K18-hACE2 transgenic mice against SARS-CoV-2 challenge. Mice were injected intraperitoneally with 5 mg/kg of either WT-VHH/12GS-WT-Fc (n = 17) or control M2e-VHH-Fc (directed against the matrix protein 2 ectodomain of influenza A; n = 10). Seven hours later, the mice were challenged with 8 x 10 3 PFU of a clinical SARS-CoV-2 isolate. ( a ) Body weight (symbols represent means ± SD, p

    Techniques Used: Activity Assay, Variant Assay, Transgenic Assay, Mouse Assay, Injection

    VHH72_S56A-Fc constructs have increased affinity and SARS-CoV-2 neutralizing activity. a. Binding affinity of VHH72-Fc variants to immobilized mouse Fc-fused SARS-CoV-2 RBD (RBD-mFc). Apparent kinetics of the 2:2 interaction is based on a global 1:1 fit of the replicate (n = 2) data; values are the averages of replicates. b. Binding of the indicated VHH72-Fc constructs (see Supplementary Table 2 for a description) to coated SARS-CoV-2 spike determined by ELISA (data points are mean ± SD; n=3). c. Binding of the indicated VHH72-Fc constructs to cell surface expressed SARS-CoV-2 spike determined by flow cytometry. The graph shows the mean (n=2) ratio of the MFI of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells. d. Inhibition of ACE-2/RBD interaction determined by AlphaLISA (amplified luminescent proximity homogeneous assay). Biotinylated SARS-CoV-2 RBD was loaded on streptavidin coated Alpha Donor beads and human ACE-2-mFc protein was captured on anti-mouse IgG acceptor beads. Interference of the donor-acceptor bead interaction was assessed for serial dilutions of the indicated VHH-Fc constructs. Graph pad Prism was used for curve fitting and IC 50 determination of triplicate measurements. e . Dose-dependent inhibition of SARS-CoV-2 RBD binding to the surface of VeroE6 cells in the presence of the indicated VHH72-Fc constructs as determined by flow cytometry. The graph shows the mean (n=2 ± SD) percentage of cells that bind RBD. f. A SARS-CoV-2 plaque reduction neutralization assay was performed with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs. Thirtysix hours after infection, the cells fixed with 3.7% paraformaldehyde and stained with 0.5% crystal violet. Data points in the graph represent the number of plaques and are representative of one experiment that was repeated once. g-h . humVHH_S56A/LALAPG-Fc and (humVHH) 2 /WT-Fc at 20 mg/kg protect hamsters against SARS-CoV-2 challenge. Hamsters were intraperitoneally injected with 20 mg/kg of palivizumab, humVHH_S56A/LALAPG-Fc or (humVHH) 2 /WT-Fc and challenged the next day with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. ( g ). Infectious virus in lungs and ( h ) viral RNA in lungs, ileum and stool determined on day 4 after the challenge. ( i ) Severity score of dilated bronchi on day 4 after the challenge.
    Figure Legend Snippet: VHH72_S56A-Fc constructs have increased affinity and SARS-CoV-2 neutralizing activity. a. Binding affinity of VHH72-Fc variants to immobilized mouse Fc-fused SARS-CoV-2 RBD (RBD-mFc). Apparent kinetics of the 2:2 interaction is based on a global 1:1 fit of the replicate (n = 2) data; values are the averages of replicates. b. Binding of the indicated VHH72-Fc constructs (see Supplementary Table 2 for a description) to coated SARS-CoV-2 spike determined by ELISA (data points are mean ± SD; n=3). c. Binding of the indicated VHH72-Fc constructs to cell surface expressed SARS-CoV-2 spike determined by flow cytometry. The graph shows the mean (n=2) ratio of the MFI of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells. d. Inhibition of ACE-2/RBD interaction determined by AlphaLISA (amplified luminescent proximity homogeneous assay). Biotinylated SARS-CoV-2 RBD was loaded on streptavidin coated Alpha Donor beads and human ACE-2-mFc protein was captured on anti-mouse IgG acceptor beads. Interference of the donor-acceptor bead interaction was assessed for serial dilutions of the indicated VHH-Fc constructs. Graph pad Prism was used for curve fitting and IC 50 determination of triplicate measurements. e . Dose-dependent inhibition of SARS-CoV-2 RBD binding to the surface of VeroE6 cells in the presence of the indicated VHH72-Fc constructs as determined by flow cytometry. The graph shows the mean (n=2 ± SD) percentage of cells that bind RBD. f. A SARS-CoV-2 plaque reduction neutralization assay was performed with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs. Thirtysix hours after infection, the cells fixed with 3.7% paraformaldehyde and stained with 0.5% crystal violet. Data points in the graph represent the number of plaques and are representative of one experiment that was repeated once. g-h . humVHH_S56A/LALAPG-Fc and (humVHH) 2 /WT-Fc at 20 mg/kg protect hamsters against SARS-CoV-2 challenge. Hamsters were intraperitoneally injected with 20 mg/kg of palivizumab, humVHH_S56A/LALAPG-Fc or (humVHH) 2 /WT-Fc and challenged the next day with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. ( g ). Infectious virus in lungs and ( h ) viral RNA in lungs, ileum and stool determined on day 4 after the challenge. ( i ) Severity score of dilated bronchi on day 4 after the challenge.

    Techniques Used: Construct, Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Transfection, Inhibition, Amplification, Neutralization, Infection, Staining, Injection

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    Article Title: A Human DPP4-Knockin Mouse’s Susceptibility to Infection by Authentic and Pseudotyped MERS-CoV
    Article Snippet: RNA was extracted from spleen cells and used to construct a phage-displayed antibody library. .. Recombinant RBD protein was used to screen antibodies. ..

    other:

    Article Title: Testing of the inhibitory effects of loratadine and desloratadine on SARS-CoV-2 spike pseudotyped virus viropexis
    Article Snippet: The copy number of pseudotyped virus was 1010 virus copies/mL, and the content of SARS-CoV-2 Spike protein was 860 ng/mL.

    Inhibition:

    Article Title: Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters
    Article Snippet: The mixtures containing RBD and VHHs or VHH-Fc fusions were added to the cells and incubated for 1 h. Subsequently, the cells were washed 3 times with PBS containing 0.5% BSA and stained with an AF647 conjugated donkey anti-mouse IgG antibody (Invitrogen) for 1 h. Following three additional washes with PBS containing 0.5% BSA, the cells were analyzed by flow cytometry using a BD LSRII flow cytometer (BD Biosciences). .. Inhibition of ACE-2/RBD interaction by AlphaLISA immunoassay Dose-dependent inhibition of the interaction of SARS-CoV-2 RBD protein with the ACE-2 receptor was assessed in a competition AlphaLISA (amplified luminescent proximity homogeneous assay). .. In brief, 2019-nCoV S protein RBD that was biotinylated through an Avi-tag (AcroBiosystems, Cat nr.

    Amplification:

    Article Title: Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters
    Article Snippet: The mixtures containing RBD and VHHs or VHH-Fc fusions were added to the cells and incubated for 1 h. Subsequently, the cells were washed 3 times with PBS containing 0.5% BSA and stained with an AF647 conjugated donkey anti-mouse IgG antibody (Invitrogen) for 1 h. Following three additional washes with PBS containing 0.5% BSA, the cells were analyzed by flow cytometry using a BD LSRII flow cytometer (BD Biosciences). .. Inhibition of ACE-2/RBD interaction by AlphaLISA immunoassay Dose-dependent inhibition of the interaction of SARS-CoV-2 RBD protein with the ACE-2 receptor was assessed in a competition AlphaLISA (amplified luminescent proximity homogeneous assay). .. In brief, 2019-nCoV S protein RBD that was biotinylated through an Avi-tag (AcroBiosystems, Cat nr.

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    Sino Biological sars cov 2 rbd protein
    HumVHH_S56A/LALA-Fc/Gen2 neutralizes <t>SARS-CoV-2</t> variants of concern. a . Surface view of SARS-CoV-2 RBD (grey) with VHH72 (green cartoon, bottom) and the N-terminal helixes of ACE2 (blue cartoon, top). The RBD-residues K417, N439, E484 and N501 (orange) are indicated. b. Binding of humVHH_S56A/LALA-Fc/Gen2 (left), CB6 (middle) and palivizumab (right) to SARS-CoV-1 spike with the RBD replaced by WT, N439K, K417N, E484K, N501Y or (K417N + E484K + N501Y) RBD of SARS-CoV-2, expressed on the surface of 293T cells. Data points represent the ratio of the mean fluorescence intensity (MFI) of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells, as determined by flow cytometry. c . SARS-CoV-2 plaque reduction neutralization assay with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs using BetaCov/Belgium/GHB-03021/2020, B1.1.7, or B.1.351 variant viruses.
    Sars Cov 2 Rbd Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, 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 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 rbd protein - by Bioz Stars, 2021-05
    96/100 stars
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    HumVHH_S56A/LALA-Fc/Gen2 neutralizes SARS-CoV-2 variants of concern. a . Surface view of SARS-CoV-2 RBD (grey) with VHH72 (green cartoon, bottom) and the N-terminal helixes of ACE2 (blue cartoon, top). The RBD-residues K417, N439, E484 and N501 (orange) are indicated. b. Binding of humVHH_S56A/LALA-Fc/Gen2 (left), CB6 (middle) and palivizumab (right) to SARS-CoV-1 spike with the RBD replaced by WT, N439K, K417N, E484K, N501Y or (K417N + E484K + N501Y) RBD of SARS-CoV-2, expressed on the surface of 293T cells. Data points represent the ratio of the mean fluorescence intensity (MFI) of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells, as determined by flow cytometry. c . SARS-CoV-2 plaque reduction neutralization assay with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs using BetaCov/Belgium/GHB-03021/2020, B1.1.7, or B.1.351 variant viruses.

    Journal: bioRxiv

    Article Title: Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters

    doi: 10.1101/2021.03.08.433449

    Figure Lengend Snippet: HumVHH_S56A/LALA-Fc/Gen2 neutralizes SARS-CoV-2 variants of concern. a . Surface view of SARS-CoV-2 RBD (grey) with VHH72 (green cartoon, bottom) and the N-terminal helixes of ACE2 (blue cartoon, top). The RBD-residues K417, N439, E484 and N501 (orange) are indicated. b. Binding of humVHH_S56A/LALA-Fc/Gen2 (left), CB6 (middle) and palivizumab (right) to SARS-CoV-1 spike with the RBD replaced by WT, N439K, K417N, E484K, N501Y or (K417N + E484K + N501Y) RBD of SARS-CoV-2, expressed on the surface of 293T cells. Data points represent the ratio of the mean fluorescence intensity (MFI) of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells, as determined by flow cytometry. c . SARS-CoV-2 plaque reduction neutralization assay with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs using BetaCov/Belgium/GHB-03021/2020, B1.1.7, or B.1.351 variant viruses.

    Article Snippet: Inhibition of ACE-2/RBD interaction by AlphaLISA immunoassay Dose-dependent inhibition of the interaction of SARS-CoV-2 RBD protein with the ACE-2 receptor was assessed in a competition AlphaLISA (amplified luminescent proximity homogeneous assay).

    Techniques: Binding Assay, Fluorescence, Transfection, Flow Cytometry, Neutralization, Construct, Variant Assay

    Therapeutic administration of VHH72-Fc constructs restricts SARS-CoV-2 virus replication in Syrian hamsters. a-c. Hamsters were challenged with 1×10 4 PFU of BetaCoV/Munich/BavPat1/2020 and 4 hours later injected intraperitoneally with 20, 7 or 2 mg/kg of bivalent humVHH_S56A/LALAPG-Fc/Gen2 or tetravalent (humVHH_S56A) 2 /LALAPG-Fc/Gen2. The negative control group was treated with 20 mg/kg of palivizumab, injected 4 hours after the challenge infection; hamsters in a prophylactic control group received 20 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day before the challenge. ( a ) Lung virus loads, ( b ) lung viral RNA copies, and ( c ) gross lung pathology determined on day 4 after infection. d,e . Hamsters received an intraperitoneal injection of 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day prior to challenge or were treated by intraperitoneal injection of 1 or 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 or (humVHH_S56A) 2 /LALAPG-Fc/Gen2 19h after infection with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Seven mg/kg of palivizumab was used as a negative control treatment. ( d ) Virus load and ( e ) viral RNA levels in the lungs on day 4 after challenge. f,g . Hamsters were treated with 4mg/kg of palivizumab, humVHH_S56A/LALA-Fc/Gen2 or humVHH/LALA-Fc/Gen2 injected intraperitoneally 24h after challenge with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Viral RNA and infectious virus were determined in lung tissue on day 4 after infection. Data were analyzed with the Mann-Whitney U-test using GraphPad Prism software. *, p

    Journal: bioRxiv

    Article Title: Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters

    doi: 10.1101/2021.03.08.433449

    Figure Lengend Snippet: Therapeutic administration of VHH72-Fc constructs restricts SARS-CoV-2 virus replication in Syrian hamsters. a-c. Hamsters were challenged with 1×10 4 PFU of BetaCoV/Munich/BavPat1/2020 and 4 hours later injected intraperitoneally with 20, 7 or 2 mg/kg of bivalent humVHH_S56A/LALAPG-Fc/Gen2 or tetravalent (humVHH_S56A) 2 /LALAPG-Fc/Gen2. The negative control group was treated with 20 mg/kg of palivizumab, injected 4 hours after the challenge infection; hamsters in a prophylactic control group received 20 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day before the challenge. ( a ) Lung virus loads, ( b ) lung viral RNA copies, and ( c ) gross lung pathology determined on day 4 after infection. d,e . Hamsters received an intraperitoneal injection of 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 one day prior to challenge or were treated by intraperitoneal injection of 1 or 7 mg/kg of humVHH_S56A/LALAPG-Fc/Gen2 or (humVHH_S56A) 2 /LALAPG-Fc/Gen2 19h after infection with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Seven mg/kg of palivizumab was used as a negative control treatment. ( d ) Virus load and ( e ) viral RNA levels in the lungs on day 4 after challenge. f,g . Hamsters were treated with 4mg/kg of palivizumab, humVHH_S56A/LALA-Fc/Gen2 or humVHH/LALA-Fc/Gen2 injected intraperitoneally 24h after challenge with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. Viral RNA and infectious virus were determined in lung tissue on day 4 after infection. Data were analyzed with the Mann-Whitney U-test using GraphPad Prism software. *, p

    Article Snippet: Inhibition of ACE-2/RBD interaction by AlphaLISA immunoassay Dose-dependent inhibition of the interaction of SARS-CoV-2 RBD protein with the ACE-2 receptor was assessed in a competition AlphaLISA (amplified luminescent proximity homogeneous assay).

    Techniques: Construct, Injection, Negative Control, Infection, MANN-WHITNEY, Software

    Enhanced affinity and neutralizing activity of computationally predicted VHH72 variant. a-c. Prototype WT-VHH/12GS-WT-Fc protects K18-hACE2 transgenic mice against SARS-CoV-2 challenge. Mice were injected intraperitoneally with 5 mg/kg of either WT-VHH/12GS-WT-Fc (n = 17) or control M2e-VHH-Fc (directed against the matrix protein 2 ectodomain of influenza A; n = 10). Seven hours later, the mice were challenged with 8 x 10 3 PFU of a clinical SARS-CoV-2 isolate. ( a ) Body weight (symbols represent means ± SD, p

    Journal: bioRxiv

    Article Title: Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters

    doi: 10.1101/2021.03.08.433449

    Figure Lengend Snippet: Enhanced affinity and neutralizing activity of computationally predicted VHH72 variant. a-c. Prototype WT-VHH/12GS-WT-Fc protects K18-hACE2 transgenic mice against SARS-CoV-2 challenge. Mice were injected intraperitoneally with 5 mg/kg of either WT-VHH/12GS-WT-Fc (n = 17) or control M2e-VHH-Fc (directed against the matrix protein 2 ectodomain of influenza A; n = 10). Seven hours later, the mice were challenged with 8 x 10 3 PFU of a clinical SARS-CoV-2 isolate. ( a ) Body weight (symbols represent means ± SD, p

    Article Snippet: Inhibition of ACE-2/RBD interaction by AlphaLISA immunoassay Dose-dependent inhibition of the interaction of SARS-CoV-2 RBD protein with the ACE-2 receptor was assessed in a competition AlphaLISA (amplified luminescent proximity homogeneous assay).

    Techniques: Activity Assay, Variant Assay, Transgenic Assay, Mouse Assay, Injection

    VHH72_S56A-Fc constructs have increased affinity and SARS-CoV-2 neutralizing activity. a. Binding affinity of VHH72-Fc variants to immobilized mouse Fc-fused SARS-CoV-2 RBD (RBD-mFc). Apparent kinetics of the 2:2 interaction is based on a global 1:1 fit of the replicate (n = 2) data; values are the averages of replicates. b. Binding of the indicated VHH72-Fc constructs (see Supplementary Table 2 for a description) to coated SARS-CoV-2 spike determined by ELISA (data points are mean ± SD; n=3). c. Binding of the indicated VHH72-Fc constructs to cell surface expressed SARS-CoV-2 spike determined by flow cytometry. The graph shows the mean (n=2) ratio of the MFI of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells. d. Inhibition of ACE-2/RBD interaction determined by AlphaLISA (amplified luminescent proximity homogeneous assay). Biotinylated SARS-CoV-2 RBD was loaded on streptavidin coated Alpha Donor beads and human ACE-2-mFc protein was captured on anti-mouse IgG acceptor beads. Interference of the donor-acceptor bead interaction was assessed for serial dilutions of the indicated VHH-Fc constructs. Graph pad Prism was used for curve fitting and IC 50 determination of triplicate measurements. e . Dose-dependent inhibition of SARS-CoV-2 RBD binding to the surface of VeroE6 cells in the presence of the indicated VHH72-Fc constructs as determined by flow cytometry. The graph shows the mean (n=2 ± SD) percentage of cells that bind RBD. f. A SARS-CoV-2 plaque reduction neutralization assay was performed with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs. Thirtysix hours after infection, the cells fixed with 3.7% paraformaldehyde and stained with 0.5% crystal violet. Data points in the graph represent the number of plaques and are representative of one experiment that was repeated once. g-h . humVHH_S56A/LALAPG-Fc and (humVHH) 2 /WT-Fc at 20 mg/kg protect hamsters against SARS-CoV-2 challenge. Hamsters were intraperitoneally injected with 20 mg/kg of palivizumab, humVHH_S56A/LALAPG-Fc or (humVHH) 2 /WT-Fc and challenged the next day with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. ( g ). Infectious virus in lungs and ( h ) viral RNA in lungs, ileum and stool determined on day 4 after the challenge. ( i ) Severity score of dilated bronchi on day 4 after the challenge.

    Journal: bioRxiv

    Article Title: Drug development of an affinity enhanced, broadly neutralizing heavy chain-only antibody that restricts SARS-CoV-2 in hamsters

    doi: 10.1101/2021.03.08.433449

    Figure Lengend Snippet: VHH72_S56A-Fc constructs have increased affinity and SARS-CoV-2 neutralizing activity. a. Binding affinity of VHH72-Fc variants to immobilized mouse Fc-fused SARS-CoV-2 RBD (RBD-mFc). Apparent kinetics of the 2:2 interaction is based on a global 1:1 fit of the replicate (n = 2) data; values are the averages of replicates. b. Binding of the indicated VHH72-Fc constructs (see Supplementary Table 2 for a description) to coated SARS-CoV-2 spike determined by ELISA (data points are mean ± SD; n=3). c. Binding of the indicated VHH72-Fc constructs to cell surface expressed SARS-CoV-2 spike determined by flow cytometry. The graph shows the mean (n=2) ratio of the MFI of transfected (GFP + ) cells over the MFI of non-transfected (GFP - ) cells. d. Inhibition of ACE-2/RBD interaction determined by AlphaLISA (amplified luminescent proximity homogeneous assay). Biotinylated SARS-CoV-2 RBD was loaded on streptavidin coated Alpha Donor beads and human ACE-2-mFc protein was captured on anti-mouse IgG acceptor beads. Interference of the donor-acceptor bead interaction was assessed for serial dilutions of the indicated VHH-Fc constructs. Graph pad Prism was used for curve fitting and IC 50 determination of triplicate measurements. e . Dose-dependent inhibition of SARS-CoV-2 RBD binding to the surface of VeroE6 cells in the presence of the indicated VHH72-Fc constructs as determined by flow cytometry. The graph shows the mean (n=2 ± SD) percentage of cells that bind RBD. f. A SARS-CoV-2 plaque reduction neutralization assay was performed with 3 fold serial dilutions of the indicated VHH-Fc fusion constructs. Thirtysix hours after infection, the cells fixed with 3.7% paraformaldehyde and stained with 0.5% crystal violet. Data points in the graph represent the number of plaques and are representative of one experiment that was repeated once. g-h . humVHH_S56A/LALAPG-Fc and (humVHH) 2 /WT-Fc at 20 mg/kg protect hamsters against SARS-CoV-2 challenge. Hamsters were intraperitoneally injected with 20 mg/kg of palivizumab, humVHH_S56A/LALAPG-Fc or (humVHH) 2 /WT-Fc and challenged the next day with 2×10 6 PFU of passage 6 BetaCov/Belgium/GHB-03021/2020. ( g ). Infectious virus in lungs and ( h ) viral RNA in lungs, ileum and stool determined on day 4 after the challenge. ( i ) Severity score of dilated bronchi on day 4 after the challenge.

    Article Snippet: Inhibition of ACE-2/RBD interaction by AlphaLISA immunoassay Dose-dependent inhibition of the interaction of SARS-CoV-2 RBD protein with the ACE-2 receptor was assessed in a competition AlphaLISA (amplified luminescent proximity homogeneous assay).

    Techniques: Construct, Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Transfection, Inhibition, Amplification, Neutralization, Infection, Staining, Injection