spike protein of sars cov 2  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike RBD AVI His Recombinant Protein Biotinylated
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike RBD AVI His Recombinant Protein YP 009724390 1 Arg319 Phe541 was expressed with a c terminal polyhistidine tagged AVI tag at the C terminus The purified protein was biotinylated in vitro
    Catalog Number:
    40592-V27H-B
    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
    Buy from Supplier


    Structured Review

    Sino Biological spike protein of sars cov 2
    Structural details at the interface between the three compounds (EP, PEP, and MEP) and human ACE2 (PDB ID: 6M0J). Amino acid residues in green present the same ACE2-binding sites with <t>SARS-CoV-2</t> RBD reported. Amino acid residues in purple present the different ACE2-binding sites with SARS-CoV-2 RBD
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike RBD AVI His Recombinant Protein YP 009724390 1 Arg319 Phe541 was expressed with a c terminal polyhistidine tagged AVI tag at the C terminus The purified protein was biotinylated in vitro
    https://www.bioz.com/result/spike protein of sars cov 2/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    spike protein of sars cov 2 - by Bioz Stars, 2021-04
    96/100 stars

    Images

    1) Product Images from "Screening and evaluation of anti-SARS-CoV-2 components from Ephedra sinica by ACE2/CMC-HPLC-IT-TOF-MS approach"

    Article Title: Screening and evaluation of anti-SARS-CoV-2 components from Ephedra sinica by ACE2/CMC-HPLC-IT-TOF-MS approach

    Journal: Analytical and Bioanalytical Chemistry

    doi: 10.1007/s00216-021-03233-7

    Structural details at the interface between the three compounds (EP, PEP, and MEP) and human ACE2 (PDB ID: 6M0J). Amino acid residues in green present the same ACE2-binding sites with SARS-CoV-2 RBD reported. Amino acid residues in purple present the different ACE2-binding sites with SARS-CoV-2 RBD
    Figure Legend Snippet: Structural details at the interface between the three compounds (EP, PEP, and MEP) and human ACE2 (PDB ID: 6M0J). Amino acid residues in green present the same ACE2-binding sites with SARS-CoV-2 RBD reported. Amino acid residues in purple present the different ACE2-binding sites with SARS-CoV-2 RBD

    Techniques Used: Binding Assay

    Effect of EP, PEP and MEP on the entrance of SARS-CoV-2 spike pseudovirus into ACE2 h cells. Inset is the graph of EP. Data are presented as mean ± SD. * P ˂ 0.05, ** P ˂ 0.01, *** P ˂ 0.001 compared with group Control
    Figure Legend Snippet: Effect of EP, PEP and MEP on the entrance of SARS-CoV-2 spike pseudovirus into ACE2 h cells. Inset is the graph of EP. Data are presented as mean ± SD. * P ˂ 0.05, ** P ˂ 0.01, *** P ˂ 0.001 compared with group Control

    Techniques Used:

    Measurement of binding affinities between the three compounds (EP, PEP and MEP) and human ACE2 (left column) or SARS-CoV-2 RBD (right column) by surface plasmon resonance assay. Purified recombinant SARS-CoV-2 RBD or recombinant human ACE2 were covalently immobilized to the sensor chip via their amine groups, and EPs flowed by. Here EPs was diluted to different concentrations (from 12.5 to 100 μM) before being injected. Data are shown as different colored lines
    Figure Legend Snippet: Measurement of binding affinities between the three compounds (EP, PEP and MEP) and human ACE2 (left column) or SARS-CoV-2 RBD (right column) by surface plasmon resonance assay. Purified recombinant SARS-CoV-2 RBD or recombinant human ACE2 were covalently immobilized to the sensor chip via their amine groups, and EPs flowed by. Here EPs was diluted to different concentrations (from 12.5 to 100 μM) before being injected. Data are shown as different colored lines

    Techniques Used: Binding Assay, SPR Assay, Purification, Recombinant, Chromatin Immunoprecipitation, Injection

    2) Product Images from "A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation"

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2021.637654

    W4P-RBD potentiates functional T cells specific to SARS-CoV-2 S1 proteins. C57BL/6 mice were intramuscularly injected with W-RBD, W4P-RBD (50 μ g /mouse), or mock, and the spleens were collected 5 weeks post-vaccination for analysis by flow cytometry. (A,B) Splenocytes were incubated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 24 h and stained to detect IFNγ-producing CD8 + T cells and CD4 + T cells. (C) Correlation between RBD-specific IgG in serum and the S1-specific T-cell population in splenocytes. Significance differences (* P
    Figure Legend Snippet: W4P-RBD potentiates functional T cells specific to SARS-CoV-2 S1 proteins. C57BL/6 mice were intramuscularly injected with W-RBD, W4P-RBD (50 μ g /mouse), or mock, and the spleens were collected 5 weeks post-vaccination for analysis by flow cytometry. (A,B) Splenocytes were incubated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 24 h and stained to detect IFNγ-producing CD8 + T cells and CD4 + T cells. (C) Correlation between RBD-specific IgG in serum and the S1-specific T-cell population in splenocytes. Significance differences (* P

    Techniques Used: Functional Assay, Mouse Assay, Injection, Flow Cytometry, Incubation, Staining

    Schematic representation of W4P-RBD as a vaccine candidate against SARS-CoV-2. A novel platform of N-terminal addition of HBV W4P preS1 33-bp sequences for a DNA vaccine against SARS-CoV-2 were developed. The W4P-RBD led to enhanced both humoral and cell-mediated immune response against SARS-CoV-2 in vaccinated mice, demonstrating its feasibility as a DNA vaccine to protect against SARS-CoV-2.
    Figure Legend Snippet: Schematic representation of W4P-RBD as a vaccine candidate against SARS-CoV-2. A novel platform of N-terminal addition of HBV W4P preS1 33-bp sequences for a DNA vaccine against SARS-CoV-2 were developed. The W4P-RBD led to enhanced both humoral and cell-mediated immune response against SARS-CoV-2 in vaccinated mice, demonstrating its feasibility as a DNA vaccine to protect against SARS-CoV-2.

    Techniques Used: Mouse Assay

    W4P-RBD elicits RBD-specific antibody responses in serum and induces potent neutralizing activity against pseudotyped SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. (A,C) Antibody responses in serum specific to SARS-CoV-2 RBD proteins were detected by ELISA. (B) Serum at 5 weeks after the last immunization was assessed using different dilution factors for IgG against the SARS-CoV-2 RBD protein using ELISA. (D) The 50% neutralizing antibody titer (NT 50 ) was calculated using the SARS-CoV-2 pseudovirus neutralization assay in Calu-3 cells. (E) Correlation between SARS-CoV-2 RBD-specific IgG and pseudotyped SARS-CoV-2 neutralization titers for immunized mice. Significance differences (* P
    Figure Legend Snippet: W4P-RBD elicits RBD-specific antibody responses in serum and induces potent neutralizing activity against pseudotyped SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. (A,C) Antibody responses in serum specific to SARS-CoV-2 RBD proteins were detected by ELISA. (B) Serum at 5 weeks after the last immunization was assessed using different dilution factors for IgG against the SARS-CoV-2 RBD protein using ELISA. (D) The 50% neutralizing antibody titer (NT 50 ) was calculated using the SARS-CoV-2 pseudovirus neutralization assay in Calu-3 cells. (E) Correlation between SARS-CoV-2 RBD-specific IgG and pseudotyped SARS-CoV-2 neutralization titers for immunized mice. Significance differences (* P

    Techniques Used: Activity Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Neutralization

    W4P-RBD exerts potent neutralizing activity against live SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) A 50% plaque reduction neutralizing antibody (PRNT 50 ) titer against live SARS-CoV-2 was calculated against SARS-CoV-2 infection in Vero E6 cells. (B) Reduction in plaque formation in Vero E6 cells infected with SARS-CoV-2. (C) Correlation between SARS-CoV-2 RBD-specific IgG and SARS-CoV-2 neutralization titers in immunized mice. Significance differences (** P
    Figure Legend Snippet: W4P-RBD exerts potent neutralizing activity against live SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) A 50% plaque reduction neutralizing antibody (PRNT 50 ) titer against live SARS-CoV-2 was calculated against SARS-CoV-2 infection in Vero E6 cells. (B) Reduction in plaque formation in Vero E6 cells infected with SARS-CoV-2. (C) Correlation between SARS-CoV-2 RBD-specific IgG and SARS-CoV-2 neutralization titers in immunized mice. Significance differences (** P

    Techniques Used: Activity Assay, Mouse Assay, Incubation, Neutralization, Plaque Reduction Neutralization Test, Infection

    The W4P-RBD vaccine exerts potent neutralizing activity against live SARS-CoV-2 and inhibits viral infection and replication of SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) The neutralization efficacy of serum from immunized mice against live SARS-CoV-2 RNA copy number in Vero E6 cells was determined by qRT-PCR. (B) The neutralization efficacy of the diluted serum against live SARS-CoV-2 in Vero E6 cells was determined by Western blotting. (C) Pooled serum (diluted 1:500) from each mouse group was tested in the neutralization assay against live SARS-CoV-2. Representative images (40-fold magnification) show live SARS-CoV-2-infected cells after neutralization in each group. Significance differences (* P
    Figure Legend Snippet: The W4P-RBD vaccine exerts potent neutralizing activity against live SARS-CoV-2 and inhibits viral infection and replication of SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) The neutralization efficacy of serum from immunized mice against live SARS-CoV-2 RNA copy number in Vero E6 cells was determined by qRT-PCR. (B) The neutralization efficacy of the diluted serum against live SARS-CoV-2 in Vero E6 cells was determined by Western blotting. (C) Pooled serum (diluted 1:500) from each mouse group was tested in the neutralization assay against live SARS-CoV-2. Representative images (40-fold magnification) show live SARS-CoV-2-infected cells after neutralization in each group. Significance differences (* P

    Techniques Used: Activity Assay, Infection, Mouse Assay, Incubation, Neutralization, Quantitative RT-PCR, Western Blot

    W4P-RBD induces proinflammatory cytokine production in S1-stimulated splenocytes. Splenocytes from immunized mice were stimulated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 5 days. The cytokine production of (A) TNFα, (B) IFNγ, (C) IL-12p40, (D) IFNβ, (E) IL-6, and (F) IL-2 from splenocytes was detected by ELISA. Significance differences (* P
    Figure Legend Snippet: W4P-RBD induces proinflammatory cytokine production in S1-stimulated splenocytes. Splenocytes from immunized mice were stimulated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 5 days. The cytokine production of (A) TNFα, (B) IFNγ, (C) IL-12p40, (D) IFNβ, (E) IL-6, and (F) IL-2 from splenocytes was detected by ELISA. Significance differences (* P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Construction of the HBV W4P preS1-fused pcDNA3.3-RBD plasmid (W4P-RBD) as a candidate for SARS-CoV-2. (A) Design of pcDNA3.3-RBD and pcDNA3.3-W4P-RBD. The W4P region comprises 33 bp from the first site of the preS1 region of the HBV genome and encodes 11 amino acids. (B) The protein expression of SARS-CoV-2 RBD and W4P-conjugated RBD was detected by the Western blot assay. pcDNA3.3-RBD, pcDNA3.3-W4P-RBD, and empty pcDNA3.3 were transfected into Vero E6, Huh7, and 293T cells, and cell lysates were collected 48 h post transfection to detect protein expression. (C) The mRNA expression levels of IL-6 and in pcDNA3.3-transfected cells were detected by qRT-PCR. Significance differences (* P
    Figure Legend Snippet: Construction of the HBV W4P preS1-fused pcDNA3.3-RBD plasmid (W4P-RBD) as a candidate for SARS-CoV-2. (A) Design of pcDNA3.3-RBD and pcDNA3.3-W4P-RBD. The W4P region comprises 33 bp from the first site of the preS1 region of the HBV genome and encodes 11 amino acids. (B) The protein expression of SARS-CoV-2 RBD and W4P-conjugated RBD was detected by the Western blot assay. pcDNA3.3-RBD, pcDNA3.3-W4P-RBD, and empty pcDNA3.3 were transfected into Vero E6, Huh7, and 293T cells, and cell lysates were collected 48 h post transfection to detect protein expression. (C) The mRNA expression levels of IL-6 and in pcDNA3.3-transfected cells were detected by qRT-PCR. Significance differences (* P

    Techniques Used: Plasmid Preparation, Expressing, Western Blot, Transfection, Quantitative RT-PCR

    3) Product Images from "Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals"

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals

    Journal: Science Translational Medicine

    doi: 10.1126/scitranslmed.abd6990

    Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.
    Figure Legend Snippet: Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.

    Techniques Used: Isolation, Binding Assay, Clone Assay, Incubation, Infection, Recombinant, Enzyme-linked Immunosorbent Assay

    Titrations of serum IgG by ELISAs specific to SARS-CoV-2. Plasma samples from 17 patients with SARS-CoV-2 were diluted (1:100) and added to plates coated with recombinant SARS-CoV-2 ( A ) N, ( B ) S, ( C ) S1, ( D ) S2, which was fused to a polyhistidine (HIS) tag, or ( E ) RBD protein, which was fused to a human hCκ domain. The amount of bound IgG was determined using anti-human IgG (Fc-specific) antibody, and ABTS (2,2’-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt) was used as the substrate. All experiments were performed in duplicate, and the data are presented as the means ± SD.
    Figure Legend Snippet: Titrations of serum IgG by ELISAs specific to SARS-CoV-2. Plasma samples from 17 patients with SARS-CoV-2 were diluted (1:100) and added to plates coated with recombinant SARS-CoV-2 ( A ) N, ( B ) S, ( C ) S1, ( D ) S2, which was fused to a polyhistidine (HIS) tag, or ( E ) RBD protein, which was fused to a human hCκ domain. The amount of bound IgG was determined using anti-human IgG (Fc-specific) antibody, and ABTS (2,2’-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt) was used as the substrate. All experiments were performed in duplicate, and the data are presented as the means ± SD.

    Techniques Used: Recombinant

    Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.
    Figure Legend Snippet: Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.

    Techniques Used: Recombinant, Mutagenesis, Incubation

    4) Product Images from "Different Innate and Adaptive Immune Responses to SARS-CoV-2 Infection of Asymptomatic, Mild, and Severe Cases"

    Article Title: Different Innate and Adaptive Immune Responses to SARS-CoV-2 Infection of Asymptomatic, Mild, and Severe Cases

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2020.610300

    (A) Arbitrary units (AU) of IgG and IgA specific for the S1 domain of the SARS-CoV-2 Spike protein and concentration of RBD specific IgM were detected by ELISA at different time points. For some patients we had the opportunity to have serum samples at different time points of the disease. Data relative to all samples collected are shown (Contacts n = 51 ; Asymptomatic n = 63 ; Mild n = 31 ; Severe n = 15 ). Midlines indicate median. Statistical significances were determined using unpaired, two-tailed Mann–Whitney U -tests. *p ≤ 0.05, **p
    Figure Legend Snippet: (A) Arbitrary units (AU) of IgG and IgA specific for the S1 domain of the SARS-CoV-2 Spike protein and concentration of RBD specific IgM were detected by ELISA at different time points. For some patients we had the opportunity to have serum samples at different time points of the disease. Data relative to all samples collected are shown (Contacts n = 51 ; Asymptomatic n = 63 ; Mild n = 31 ; Severe n = 15 ). Midlines indicate median. Statistical significances were determined using unpaired, two-tailed Mann–Whitney U -tests. *p ≤ 0.05, **p

    Techniques Used: Concentration Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY

    5) Product Images from "Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals"

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals

    Journal: Science Translational Medicine

    doi: 10.1126/scitranslmed.abd6990

    Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.
    Figure Legend Snippet: Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.

    Techniques Used: Isolation, Binding Assay, Clone Assay, Incubation, Infection, Recombinant, Enzyme-linked Immunosorbent Assay

    Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.
    Figure Legend Snippet: Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.

    Techniques Used: Recombinant, Mutagenesis, Incubation

    6) Product Images from "Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals"

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals

    Journal: Science Translational Medicine

    doi: 10.1126/scitranslmed.abd6990

    Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.
    Figure Legend Snippet: Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.

    Techniques Used: Isolation, Binding Assay, Clone Assay, Incubation, Infection, Recombinant, Enzyme-linked Immunosorbent Assay

    Titrations of serum IgG by ELISAs specific to SARS-CoV-2. Plasma samples from 17 patients with SARS-CoV-2 were diluted (1:100) and added to plates coated with recombinant SARS-CoV-2 ( A ) N, ( B ) S, ( C ) S1, ( D ) S2, which was fused to a polyhistidine (HIS) tag, or ( E ) RBD protein, which was fused to a human hCκ domain. The amount of bound IgG was determined using anti-human IgG (Fc-specific) antibody, and ABTS (2,2’-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt) was used as the substrate. All experiments were performed in duplicate, and the data are presented as the means ± SD.
    Figure Legend Snippet: Titrations of serum IgG by ELISAs specific to SARS-CoV-2. Plasma samples from 17 patients with SARS-CoV-2 were diluted (1:100) and added to plates coated with recombinant SARS-CoV-2 ( A ) N, ( B ) S, ( C ) S1, ( D ) S2, which was fused to a polyhistidine (HIS) tag, or ( E ) RBD protein, which was fused to a human hCκ domain. The amount of bound IgG was determined using anti-human IgG (Fc-specific) antibody, and ABTS (2,2’-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt) was used as the substrate. All experiments were performed in duplicate, and the data are presented as the means ± SD.

    Techniques Used: Recombinant

    Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.
    Figure Legend Snippet: Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.

    Techniques Used: Recombinant, Mutagenesis, Incubation

    Related Articles

    other:

    Article Title: Membrane Nanoparticles Derived from ACE2-rich Cells Block SARS-CoV-2 Infection
    Article Snippet: Biotinylated SARS-CoV-2 RBD and D614G-S1 were obtained with a G-MM-IGT biotinylation kit (Genemore, Shanghai, CHN) was immobilized on SA biosensors at 15 μg mL−1 .

    Recombinant:

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals
    Article Snippet: Then, IgG2/4 was purified by affinity chromatography using MabSelect columns with the AKTA pure chromatography system (GE Healthcare) following the manufacturer’s protocol. .. Enzyme-linked immunosorbent assayOne hundred nanograms of each recombinant SARS-CoV-2 S (Sino Biological Inc.), S1 (Sino Biological Inc.), S1 D614G (Sino Biological Inc.), S2 (Sino Biological Inc.), nucleocapsid (N) (Sino Biological Inc.), RBD, RBD mutants, SARS-CoV RBD (Sino Biological Inc.), MERS-CoV S (Sino Biological Inc.), RBD (Sino Biological Inc.), and S2 (Sino Biological Inc.) proteins were added to microtiter plates (CoStar), in coating buffer (0.1 M sodium bicarbonate, pH 8.6). .. After incubation at 4°C overnight and blocking with 3% bovine serum albumin (BSA) in PBS, for 1 hour at 37°C, serially diluted plasma (fivefold, six dilutions, starting from 1:100) or scFv-hFc (fivefold, 12 dilutions, starting from 1000 or 500 nM) in blocking buffer was added to individual wells and incubated for 1 hour at 37°C.

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals
    Article Snippet: .. These libraries were subjected to four rounds of biopanning against the recombinant SARS-CoV-2 RBD protein (Sino Biological Inc., Beijing, China), fused to mouse Fc or hCκ, as described previously ( ). .. Briefly, 3 μg of the recombinant SARS-CoV-2 RBD protein was conjugated to 1.0 × 107 magnetic beads (Dynabeads M-270 epoxy, Invitrogen) and incubated with the scFv phage display libraries (about 1012 phages), for 2 hours at 37°C.

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals
    Article Snippet: During the first round of biopanning, the beads were washed once with 500 μl of 0.05% (v/v) Tween 20 (Sigma-Aldrich, St. Louis, MO, USA) in phosphate-buffered saline (PBST). .. For the other rounds of biopanning, 1.5 μg of recombinant SARS-CoV-2 RBD protein was conjugated to 5.0 × 106 magnetic beads, and the number of washes was increased to three. ..

    Staining:

    Article Title: Low-Dose Ad26.COV2.S Protection Against SARS-CoV-2 Challenge in Rhesus Macaques
    Article Snippet: .. B cell immunophenotypingFresh PBMCs were stained with Aqua live/dead dye for 20 min, washed with 2% FBS/DPBS buffer, and cells were suspended in 2% FBS/DPBS buffer with Fc Block (BD) for 10 min, followed by staining with monoclonal antibodies against CD45 (clone D058-1283, BUV805), CD3 (clone SP34.2, APC-Cy7), CD7 (clone M-T701, Alexa700), CD123 (clone 6H6, Alexa700), CD11c (clone 3.9, Alexa700), CD20 (clone 2H7, PE-Cy5), IgA (goat polyclonal antibodies, APC), IgG (clone G18-145, BUV737), IgM (clone G20-127, BUV396), IgD (goat polyclonal antibodies, PE), CD80 (clone L307.4, BV786), CD95 (clone DX2, BV711), CD27 (clone M-T271, BUV563), CD21 (clone B-ly4, BV605), CD14 (clone M5E2, BV570), CD138 (clone DL-101, PE-CF594), and staining with SARS-CoV-2 antigens including biotinylated SARS-CoV-2 RBD proteins (Sino Biological) and full-length SARS-CoV-2 spike proteins (Sino Biological) labeled with FITC and DyLight 405, at 4 °C for 30 min. After staining, cells were washed twice with 2% FBS/DPBS buffer, followed by incubation with BV650 streptavidin (BD Pharmingen) for 10min, then washed twice with 2% FBS/DPBS buffer. .. For intracellular staining, cells were permeabilized using Caltag Fix & Perm (Invitrogen), then stained with monoclonal antibodies against Ki67 (clone B56, PerCP-cy5.5) and IRF4 (clone 3E4, PE-Cy7).

    Blocking Assay:

    Article Title: Low-Dose Ad26.COV2.S Protection Against SARS-CoV-2 Challenge in Rhesus Macaques
    Article Snippet: .. B cell immunophenotypingFresh PBMCs were stained with Aqua live/dead dye for 20 min, washed with 2% FBS/DPBS buffer, and cells were suspended in 2% FBS/DPBS buffer with Fc Block (BD) for 10 min, followed by staining with monoclonal antibodies against CD45 (clone D058-1283, BUV805), CD3 (clone SP34.2, APC-Cy7), CD7 (clone M-T701, Alexa700), CD123 (clone 6H6, Alexa700), CD11c (clone 3.9, Alexa700), CD20 (clone 2H7, PE-Cy5), IgA (goat polyclonal antibodies, APC), IgG (clone G18-145, BUV737), IgM (clone G20-127, BUV396), IgD (goat polyclonal antibodies, PE), CD80 (clone L307.4, BV786), CD95 (clone DX2, BV711), CD27 (clone M-T271, BUV563), CD21 (clone B-ly4, BV605), CD14 (clone M5E2, BV570), CD138 (clone DL-101, PE-CF594), and staining with SARS-CoV-2 antigens including biotinylated SARS-CoV-2 RBD proteins (Sino Biological) and full-length SARS-CoV-2 spike proteins (Sino Biological) labeled with FITC and DyLight 405, at 4 °C for 30 min. After staining, cells were washed twice with 2% FBS/DPBS buffer, followed by incubation with BV650 streptavidin (BD Pharmingen) for 10min, then washed twice with 2% FBS/DPBS buffer. .. For intracellular staining, cells were permeabilized using Caltag Fix & Perm (Invitrogen), then stained with monoclonal antibodies against Ki67 (clone B56, PerCP-cy5.5) and IRF4 (clone 3E4, PE-Cy7).

    Labeling:

    Article Title: Low-Dose Ad26.COV2.S Protection Against SARS-CoV-2 Challenge in Rhesus Macaques
    Article Snippet: .. B cell immunophenotypingFresh PBMCs were stained with Aqua live/dead dye for 20 min, washed with 2% FBS/DPBS buffer, and cells were suspended in 2% FBS/DPBS buffer with Fc Block (BD) for 10 min, followed by staining with monoclonal antibodies against CD45 (clone D058-1283, BUV805), CD3 (clone SP34.2, APC-Cy7), CD7 (clone M-T701, Alexa700), CD123 (clone 6H6, Alexa700), CD11c (clone 3.9, Alexa700), CD20 (clone 2H7, PE-Cy5), IgA (goat polyclonal antibodies, APC), IgG (clone G18-145, BUV737), IgM (clone G20-127, BUV396), IgD (goat polyclonal antibodies, PE), CD80 (clone L307.4, BV786), CD95 (clone DX2, BV711), CD27 (clone M-T271, BUV563), CD21 (clone B-ly4, BV605), CD14 (clone M5E2, BV570), CD138 (clone DL-101, PE-CF594), and staining with SARS-CoV-2 antigens including biotinylated SARS-CoV-2 RBD proteins (Sino Biological) and full-length SARS-CoV-2 spike proteins (Sino Biological) labeled with FITC and DyLight 405, at 4 °C for 30 min. After staining, cells were washed twice with 2% FBS/DPBS buffer, followed by incubation with BV650 streptavidin (BD Pharmingen) for 10min, then washed twice with 2% FBS/DPBS buffer. .. For intracellular staining, cells were permeabilized using Caltag Fix & Perm (Invitrogen), then stained with monoclonal antibodies against Ki67 (clone B56, PerCP-cy5.5) and IRF4 (clone 3E4, PE-Cy7).

    Incubation:

    Article Title: Low-Dose Ad26.COV2.S Protection Against SARS-CoV-2 Challenge in Rhesus Macaques
    Article Snippet: .. B cell immunophenotypingFresh PBMCs were stained with Aqua live/dead dye for 20 min, washed with 2% FBS/DPBS buffer, and cells were suspended in 2% FBS/DPBS buffer with Fc Block (BD) for 10 min, followed by staining with monoclonal antibodies against CD45 (clone D058-1283, BUV805), CD3 (clone SP34.2, APC-Cy7), CD7 (clone M-T701, Alexa700), CD123 (clone 6H6, Alexa700), CD11c (clone 3.9, Alexa700), CD20 (clone 2H7, PE-Cy5), IgA (goat polyclonal antibodies, APC), IgG (clone G18-145, BUV737), IgM (clone G20-127, BUV396), IgD (goat polyclonal antibodies, PE), CD80 (clone L307.4, BV786), CD95 (clone DX2, BV711), CD27 (clone M-T271, BUV563), CD21 (clone B-ly4, BV605), CD14 (clone M5E2, BV570), CD138 (clone DL-101, PE-CF594), and staining with SARS-CoV-2 antigens including biotinylated SARS-CoV-2 RBD proteins (Sino Biological) and full-length SARS-CoV-2 spike proteins (Sino Biological) labeled with FITC and DyLight 405, at 4 °C for 30 min. After staining, cells were washed twice with 2% FBS/DPBS buffer, followed by incubation with BV650 streptavidin (BD Pharmingen) for 10min, then washed twice with 2% FBS/DPBS buffer. .. For intracellular staining, cells were permeabilized using Caltag Fix & Perm (Invitrogen), then stained with monoclonal antibodies against Ki67 (clone B56, PerCP-cy5.5) and IRF4 (clone 3E4, PE-Cy7).

    Purification:

    Article Title: Different Innate and Adaptive Immune Responses to SARS-CoV-2 Infection of Asymptomatic, Mild, and Severe Cases
    Article Snippet: For the detection of IgM anti RBD we developed an in-house ELISA. .. 96-well plates (Corning) were coated overnight with 1 μg/ml of purified SARS-CoV-2 RBD protein (Sino Biological). .. After washing with PBS/0.05% Tween and blocking with PBS/1% BSA, plates were incubated for 1 h at 37°C with diluted sera.

    SDS Page:

    Article Title: Screening and evaluation of anti-SARS-CoV-2 components from Ephedra sinica by ACE2/CMC-HPLC-IT-TOF-MS approach
    Article Snippet: .. The receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 (SARS-CoV-2 RBD, purity > 95% as determined by SDS-PAGE) and human ACE2 protein (purity > 95% as determined by SDS-PAGE) were supplied by Sino Biological Inc. (Beijing, China). .. ACE2h cell culture ACE2 over-expressed HEK293T cell line (ACE2h cell line) was constructed and verified by Genomeditech (Shanghai, China) and was cultured in DMEM with high glucose containing 10% PBS, 1% penicillin-streptomycin, and 4 μg/mL puromycin; cells were seeded in a flask and kept at 37 °C in a 5% CO2 incubator.

    Amplification:

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation
    Article Snippet: Design and Synthesis of pcDNA3.3-RBD and pcDNA3.3-W4P-RBD pcDNA3.3-RBD and pcDNA3.3-W4P-RBD were constructed and synthesized as follows. .. Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template. .. Genes encoding W4P-RBD were amplified using primers.

    Expressing:

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation
    Article Snippet: Design and Synthesis of pcDNA3.3-RBD and pcDNA3.3-W4P-RBD pcDNA3.3-RBD and pcDNA3.3-W4P-RBD were constructed and synthesized as follows. .. Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template. .. Genes encoding W4P-RBD were amplified using primers.

    Plasmid Preparation:

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation
    Article Snippet: Design and Synthesis of pcDNA3.3-RBD and pcDNA3.3-W4P-RBD pcDNA3.3-RBD and pcDNA3.3-W4P-RBD were constructed and synthesized as follows. .. Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template. .. Genes encoding W4P-RBD were amplified using primers.

    Magnetic Beads:

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals
    Article Snippet: During the first round of biopanning, the beads were washed once with 500 μl of 0.05% (v/v) Tween 20 (Sigma-Aldrich, St. Louis, MO, USA) in phosphate-buffered saline (PBST). .. For the other rounds of biopanning, 1.5 μg of recombinant SARS-CoV-2 RBD protein was conjugated to 5.0 × 106 magnetic beads, and the number of washes was increased to three. ..

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    Sino Biological spike protein of sars cov 2
    Structural details at the interface between the three compounds (EP, PEP, and MEP) and human ACE2 (PDB ID: 6M0J). Amino acid residues in green present the same ACE2-binding sites with <t>SARS-CoV-2</t> RBD reported. Amino acid residues in purple present the different ACE2-binding sites with SARS-CoV-2 RBD
    Spike Protein Of Sars Cov 2, 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
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    Structural details at the interface between the three compounds (EP, PEP, and MEP) and human ACE2 (PDB ID: 6M0J). Amino acid residues in green present the same ACE2-binding sites with SARS-CoV-2 RBD reported. Amino acid residues in purple present the different ACE2-binding sites with SARS-CoV-2 RBD

    Journal: Analytical and Bioanalytical Chemistry

    Article Title: Screening and evaluation of anti-SARS-CoV-2 components from Ephedra sinica by ACE2/CMC-HPLC-IT-TOF-MS approach

    doi: 10.1007/s00216-021-03233-7

    Figure Lengend Snippet: Structural details at the interface between the three compounds (EP, PEP, and MEP) and human ACE2 (PDB ID: 6M0J). Amino acid residues in green present the same ACE2-binding sites with SARS-CoV-2 RBD reported. Amino acid residues in purple present the different ACE2-binding sites with SARS-CoV-2 RBD

    Article Snippet: The receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 (SARS-CoV-2 RBD, purity > 95% as determined by SDS-PAGE) and human ACE2 protein (purity > 95% as determined by SDS-PAGE) were supplied by Sino Biological Inc. (Beijing, China).

    Techniques: Binding Assay

    Effect of EP, PEP and MEP on the entrance of SARS-CoV-2 spike pseudovirus into ACE2 h cells. Inset is the graph of EP. Data are presented as mean ± SD. * P ˂ 0.05, ** P ˂ 0.01, *** P ˂ 0.001 compared with group Control

    Journal: Analytical and Bioanalytical Chemistry

    Article Title: Screening and evaluation of anti-SARS-CoV-2 components from Ephedra sinica by ACE2/CMC-HPLC-IT-TOF-MS approach

    doi: 10.1007/s00216-021-03233-7

    Figure Lengend Snippet: Effect of EP, PEP and MEP on the entrance of SARS-CoV-2 spike pseudovirus into ACE2 h cells. Inset is the graph of EP. Data are presented as mean ± SD. * P ˂ 0.05, ** P ˂ 0.01, *** P ˂ 0.001 compared with group Control

    Article Snippet: The receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 (SARS-CoV-2 RBD, purity > 95% as determined by SDS-PAGE) and human ACE2 protein (purity > 95% as determined by SDS-PAGE) were supplied by Sino Biological Inc. (Beijing, China).

    Techniques:

    Measurement of binding affinities between the three compounds (EP, PEP and MEP) and human ACE2 (left column) or SARS-CoV-2 RBD (right column) by surface plasmon resonance assay. Purified recombinant SARS-CoV-2 RBD or recombinant human ACE2 were covalently immobilized to the sensor chip via their amine groups, and EPs flowed by. Here EPs was diluted to different concentrations (from 12.5 to 100 μM) before being injected. Data are shown as different colored lines

    Journal: Analytical and Bioanalytical Chemistry

    Article Title: Screening and evaluation of anti-SARS-CoV-2 components from Ephedra sinica by ACE2/CMC-HPLC-IT-TOF-MS approach

    doi: 10.1007/s00216-021-03233-7

    Figure Lengend Snippet: Measurement of binding affinities between the three compounds (EP, PEP and MEP) and human ACE2 (left column) or SARS-CoV-2 RBD (right column) by surface plasmon resonance assay. Purified recombinant SARS-CoV-2 RBD or recombinant human ACE2 were covalently immobilized to the sensor chip via their amine groups, and EPs flowed by. Here EPs was diluted to different concentrations (from 12.5 to 100 μM) before being injected. Data are shown as different colored lines

    Article Snippet: The receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 (SARS-CoV-2 RBD, purity > 95% as determined by SDS-PAGE) and human ACE2 protein (purity > 95% as determined by SDS-PAGE) were supplied by Sino Biological Inc. (Beijing, China).

    Techniques: Binding Assay, SPR Assay, Purification, Recombinant, Chromatin Immunoprecipitation, Injection

    W4P-RBD potentiates functional T cells specific to SARS-CoV-2 S1 proteins. C57BL/6 mice were intramuscularly injected with W-RBD, W4P-RBD (50 μ g /mouse), or mock, and the spleens were collected 5 weeks post-vaccination for analysis by flow cytometry. (A,B) Splenocytes were incubated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 24 h and stained to detect IFNγ-producing CD8 + T cells and CD4 + T cells. (C) Correlation between RBD-specific IgG in serum and the S1-specific T-cell population in splenocytes. Significance differences (* P

    Journal: Frontiers in Immunology

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    doi: 10.3389/fimmu.2021.637654

    Figure Lengend Snippet: W4P-RBD potentiates functional T cells specific to SARS-CoV-2 S1 proteins. C57BL/6 mice were intramuscularly injected with W-RBD, W4P-RBD (50 μ g /mouse), or mock, and the spleens were collected 5 weeks post-vaccination for analysis by flow cytometry. (A,B) Splenocytes were incubated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 24 h and stained to detect IFNγ-producing CD8 + T cells and CD4 + T cells. (C) Correlation between RBD-specific IgG in serum and the S1-specific T-cell population in splenocytes. Significance differences (* P

    Article Snippet: Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template.

    Techniques: Functional Assay, Mouse Assay, Injection, Flow Cytometry, Incubation, Staining

    Schematic representation of W4P-RBD as a vaccine candidate against SARS-CoV-2. A novel platform of N-terminal addition of HBV W4P preS1 33-bp sequences for a DNA vaccine against SARS-CoV-2 were developed. The W4P-RBD led to enhanced both humoral and cell-mediated immune response against SARS-CoV-2 in vaccinated mice, demonstrating its feasibility as a DNA vaccine to protect against SARS-CoV-2.

    Journal: Frontiers in Immunology

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    doi: 10.3389/fimmu.2021.637654

    Figure Lengend Snippet: Schematic representation of W4P-RBD as a vaccine candidate against SARS-CoV-2. A novel platform of N-terminal addition of HBV W4P preS1 33-bp sequences for a DNA vaccine against SARS-CoV-2 were developed. The W4P-RBD led to enhanced both humoral and cell-mediated immune response against SARS-CoV-2 in vaccinated mice, demonstrating its feasibility as a DNA vaccine to protect against SARS-CoV-2.

    Article Snippet: Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template.

    Techniques: Mouse Assay

    W4P-RBD elicits RBD-specific antibody responses in serum and induces potent neutralizing activity against pseudotyped SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. (A,C) Antibody responses in serum specific to SARS-CoV-2 RBD proteins were detected by ELISA. (B) Serum at 5 weeks after the last immunization was assessed using different dilution factors for IgG against the SARS-CoV-2 RBD protein using ELISA. (D) The 50% neutralizing antibody titer (NT 50 ) was calculated using the SARS-CoV-2 pseudovirus neutralization assay in Calu-3 cells. (E) Correlation between SARS-CoV-2 RBD-specific IgG and pseudotyped SARS-CoV-2 neutralization titers for immunized mice. Significance differences (* P

    Journal: Frontiers in Immunology

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    doi: 10.3389/fimmu.2021.637654

    Figure Lengend Snippet: W4P-RBD elicits RBD-specific antibody responses in serum and induces potent neutralizing activity against pseudotyped SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. (A,C) Antibody responses in serum specific to SARS-CoV-2 RBD proteins were detected by ELISA. (B) Serum at 5 weeks after the last immunization was assessed using different dilution factors for IgG against the SARS-CoV-2 RBD protein using ELISA. (D) The 50% neutralizing antibody titer (NT 50 ) was calculated using the SARS-CoV-2 pseudovirus neutralization assay in Calu-3 cells. (E) Correlation between SARS-CoV-2 RBD-specific IgG and pseudotyped SARS-CoV-2 neutralization titers for immunized mice. Significance differences (* P

    Article Snippet: Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template.

    Techniques: Activity Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Neutralization

    W4P-RBD exerts potent neutralizing activity against live SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) A 50% plaque reduction neutralizing antibody (PRNT 50 ) titer against live SARS-CoV-2 was calculated against SARS-CoV-2 infection in Vero E6 cells. (B) Reduction in plaque formation in Vero E6 cells infected with SARS-CoV-2. (C) Correlation between SARS-CoV-2 RBD-specific IgG and SARS-CoV-2 neutralization titers in immunized mice. Significance differences (** P

    Journal: Frontiers in Immunology

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    doi: 10.3389/fimmu.2021.637654

    Figure Lengend Snippet: W4P-RBD exerts potent neutralizing activity against live SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) A 50% plaque reduction neutralizing antibody (PRNT 50 ) titer against live SARS-CoV-2 was calculated against SARS-CoV-2 infection in Vero E6 cells. (B) Reduction in plaque formation in Vero E6 cells infected with SARS-CoV-2. (C) Correlation between SARS-CoV-2 RBD-specific IgG and SARS-CoV-2 neutralization titers in immunized mice. Significance differences (** P

    Article Snippet: Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template.

    Techniques: Activity Assay, Mouse Assay, Incubation, Neutralization, Plaque Reduction Neutralization Test, Infection

    The W4P-RBD vaccine exerts potent neutralizing activity against live SARS-CoV-2 and inhibits viral infection and replication of SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) The neutralization efficacy of serum from immunized mice against live SARS-CoV-2 RNA copy number in Vero E6 cells was determined by qRT-PCR. (B) The neutralization efficacy of the diluted serum against live SARS-CoV-2 in Vero E6 cells was determined by Western blotting. (C) Pooled serum (diluted 1:500) from each mouse group was tested in the neutralization assay against live SARS-CoV-2. Representative images (40-fold magnification) show live SARS-CoV-2-infected cells after neutralization in each group. Significance differences (* P

    Journal: Frontiers in Immunology

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    doi: 10.3389/fimmu.2021.637654

    Figure Lengend Snippet: The W4P-RBD vaccine exerts potent neutralizing activity against live SARS-CoV-2 and inhibits viral infection and replication of SARS-CoV-2. C57BL/6 mice were immunized with W-RBD, W4P-RBD (50 μ g /mouse), or empty pcDNA3.3 (Mock) three times at 1-week intervals. Serum from the immunized mice was diluted and incubated with live SARS-CoV-2 for neutralization assays. (A) The neutralization efficacy of serum from immunized mice against live SARS-CoV-2 RNA copy number in Vero E6 cells was determined by qRT-PCR. (B) The neutralization efficacy of the diluted serum against live SARS-CoV-2 in Vero E6 cells was determined by Western blotting. (C) Pooled serum (diluted 1:500) from each mouse group was tested in the neutralization assay against live SARS-CoV-2. Representative images (40-fold magnification) show live SARS-CoV-2-infected cells after neutralization in each group. Significance differences (* P

    Article Snippet: Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template.

    Techniques: Activity Assay, Infection, Mouse Assay, Incubation, Neutralization, Quantitative RT-PCR, Western Blot

    W4P-RBD induces proinflammatory cytokine production in S1-stimulated splenocytes. Splenocytes from immunized mice were stimulated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 5 days. The cytokine production of (A) TNFα, (B) IFNγ, (C) IL-12p40, (D) IFNβ, (E) IL-6, and (F) IL-2 from splenocytes was detected by ELISA. Significance differences (* P

    Journal: Frontiers in Immunology

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    doi: 10.3389/fimmu.2021.637654

    Figure Lengend Snippet: W4P-RBD induces proinflammatory cytokine production in S1-stimulated splenocytes. Splenocytes from immunized mice were stimulated with SARS-CoV-2 S1 protein (5 μ g / ml ) for 5 days. The cytokine production of (A) TNFα, (B) IFNγ, (C) IL-12p40, (D) IFNβ, (E) IL-6, and (F) IL-2 from splenocytes was detected by ELISA. Significance differences (* P

    Article Snippet: Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Construction of the HBV W4P preS1-fused pcDNA3.3-RBD plasmid (W4P-RBD) as a candidate for SARS-CoV-2. (A) Design of pcDNA3.3-RBD and pcDNA3.3-W4P-RBD. The W4P region comprises 33 bp from the first site of the preS1 region of the HBV genome and encodes 11 amino acids. (B) The protein expression of SARS-CoV-2 RBD and W4P-conjugated RBD was detected by the Western blot assay. pcDNA3.3-RBD, pcDNA3.3-W4P-RBD, and empty pcDNA3.3 were transfected into Vero E6, Huh7, and 293T cells, and cell lysates were collected 48 h post transfection to detect protein expression. (C) The mRNA expression levels of IL-6 and in pcDNA3.3-transfected cells were detected by qRT-PCR. Significance differences (* P

    Journal: Frontiers in Immunology

    Article Title: A Novel DNA Vaccine Against SARS-CoV-2 Encoding a Chimeric Protein of Its Receptor-Binding Domain (RBD) Fused to the Amino-Terminal Region of Hepatitis B Virus preS1 With a W4P Mutation

    doi: 10.3389/fimmu.2021.637654

    Figure Lengend Snippet: Construction of the HBV W4P preS1-fused pcDNA3.3-RBD plasmid (W4P-RBD) as a candidate for SARS-CoV-2. (A) Design of pcDNA3.3-RBD and pcDNA3.3-W4P-RBD. The W4P region comprises 33 bp from the first site of the preS1 region of the HBV genome and encodes 11 amino acids. (B) The protein expression of SARS-CoV-2 RBD and W4P-conjugated RBD was detected by the Western blot assay. pcDNA3.3-RBD, pcDNA3.3-W4P-RBD, and empty pcDNA3.3 were transfected into Vero E6, Huh7, and 293T cells, and cell lysates were collected 48 h post transfection to detect protein expression. (C) The mRNA expression levels of IL-6 and in pcDNA3.3-transfected cells were detected by qRT-PCR. Significance differences (* P

    Article Snippet: Briefly, genes encoding RBD (residue 319-541) of the SARS-CoV-2 spike protein were amplified using RBD primers and codon-optimized SARS-CoV-2 Spike ORF mammalian expression plasmid (VG40589; Sino Biological, CN) as the template.

    Techniques: Plasmid Preparation, Expressing, Western Blot, Transfection, Quantitative RT-PCR

    Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.

    Journal: Science Translational Medicine

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals

    doi: 10.1126/scitranslmed.abd6990

    Figure Lengend Snippet: Characteristics of the isolated nAbs, stereotypic IGH clonotypes, and RBD binding–predicted clones. ( A ) Serially diluted IgG2/4 was mixed with an equal volume of SARS-CoV-2 containing 100 TCID 50 , and the IgG2/4-virus mixture was added to Vero cells with eight repeats and incubated for 5 days. Cells infected with 100 TCID 50 of SARS-CoV-2, isotype IgG2/4 control, or without the virus were applied as positive, negative, and uninfected controls, respectively. CPE in each well was observed 5 days after infection. ( B ) Characteristics of nAbs found in patients A and E. ( C ) IGH clonotypes that are highly homologous to E-3B1 and reactive against recombinant SARS-CoV-2 S and RBD proteins. The right column shows the results of the phage ELISA. All experiments were performed in quadruplicate, and the data are presented as the means ± SD. ( D ) List of diverse Ig light chain clonotypes that can be paired with the IGH clonotypes from (B) to achieve reactivity. ( E ) J and ( F ) VJ gene usage in the IGH repertoire of patients (top) and the binding-predicted IGH clones (bottom). For the VJ gene usage heatmap, the frequency values for the IGH repertoire of all 17 patients were averaged and are displayed (top) along with those of the predicted RBD-binding IGH clones (bottom). N/A, not applicable.

    Article Snippet: For the other rounds of biopanning, 1.5 μg of recombinant SARS-CoV-2 RBD protein was conjugated to 5.0 × 106 magnetic beads, and the number of washes was increased to three.

    Techniques: Isolation, Binding Assay, Clone Assay, Incubation, Infection, Recombinant, Enzyme-linked Immunosorbent Assay

    Titrations of serum IgG by ELISAs specific to SARS-CoV-2. Plasma samples from 17 patients with SARS-CoV-2 were diluted (1:100) and added to plates coated with recombinant SARS-CoV-2 ( A ) N, ( B ) S, ( C ) S1, ( D ) S2, which was fused to a polyhistidine (HIS) tag, or ( E ) RBD protein, which was fused to a human hCκ domain. The amount of bound IgG was determined using anti-human IgG (Fc-specific) antibody, and ABTS (2,2’-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt) was used as the substrate. All experiments were performed in duplicate, and the data are presented as the means ± SD.

    Journal: Science Translational Medicine

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals

    doi: 10.1126/scitranslmed.abd6990

    Figure Lengend Snippet: Titrations of serum IgG by ELISAs specific to SARS-CoV-2. Plasma samples from 17 patients with SARS-CoV-2 were diluted (1:100) and added to plates coated with recombinant SARS-CoV-2 ( A ) N, ( B ) S, ( C ) S1, ( D ) S2, which was fused to a polyhistidine (HIS) tag, or ( E ) RBD protein, which was fused to a human hCκ domain. The amount of bound IgG was determined using anti-human IgG (Fc-specific) antibody, and ABTS (2,2’-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt) was used as the substrate. All experiments were performed in duplicate, and the data are presented as the means ± SD.

    Article Snippet: For the other rounds of biopanning, 1.5 μg of recombinant SARS-CoV-2 RBD protein was conjugated to 5.0 × 106 magnetic beads, and the number of washes was increased to three.

    Techniques: Recombinant

    Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.

    Journal: Science Translational Medicine

    Article Title: Stereotypic neutralizing VH antibodies against SARS-CoV-2 spike protein receptor binding domain in patients with COVID-19 and healthy individuals

    doi: 10.1126/scitranslmed.abd6990

    Figure Lengend Snippet: Reactivity of nAbs against recombinant SARS-CoV-2 spike mutants. Recombinant wild-type or mutant (V341I, F342L, N354D, V367F, R408I, A435S, G476S, V483A, and D614G) SARS-CoV-2 S, S1, or RBD protein–coated microtiter plates were incubated with varying concentrations of ( A ) E-3B1-hFc, ( B ) A-1H4-hFc, ( C ) A-2F1-hFc, ( D ) A-2H4-hFc, ( E ) E-3G9-hFc, and ( F ) irrelevant scFv-hFc. HRP-conjugated anti-human IgG antibody was used as the probe, and ABTS was used as the substrate. All experiments were performed in triplicate, and data are presented as the means ± SD.

    Article Snippet: For the other rounds of biopanning, 1.5 μg of recombinant SARS-CoV-2 RBD protein was conjugated to 5.0 × 106 magnetic beads, and the number of washes was increased to three.

    Techniques: Recombinant, Mutagenesis, Incubation

    (A) Arbitrary units (AU) of IgG and IgA specific for the S1 domain of the SARS-CoV-2 Spike protein and concentration of RBD specific IgM were detected by ELISA at different time points. For some patients we had the opportunity to have serum samples at different time points of the disease. Data relative to all samples collected are shown (Contacts n = 51 ; Asymptomatic n = 63 ; Mild n = 31 ; Severe n = 15 ). Midlines indicate median. Statistical significances were determined using unpaired, two-tailed Mann–Whitney U -tests. *p ≤ 0.05, **p

    Journal: Frontiers in Immunology

    Article Title: Different Innate and Adaptive Immune Responses to SARS-CoV-2 Infection of Asymptomatic, Mild, and Severe Cases

    doi: 10.3389/fimmu.2020.610300

    Figure Lengend Snippet: (A) Arbitrary units (AU) of IgG and IgA specific for the S1 domain of the SARS-CoV-2 Spike protein and concentration of RBD specific IgM were detected by ELISA at different time points. For some patients we had the opportunity to have serum samples at different time points of the disease. Data relative to all samples collected are shown (Contacts n = 51 ; Asymptomatic n = 63 ; Mild n = 31 ; Severe n = 15 ). Midlines indicate median. Statistical significances were determined using unpaired, two-tailed Mann–Whitney U -tests. *p ≤ 0.05, **p

    Article Snippet: 96-well plates (Corning) were coated overnight with 1 μg/ml of purified SARS-CoV-2 RBD protein (Sino Biological).

    Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test, MANN-WHITNEY