mouse igg1 fc domain  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike RBD mFc Recombinant Protein HPLC verified COVID 19 Spike RBD Research
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with the Fc region of mouse IgG1 at the C terminus
    Catalog Number:
    40592-V05H
    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 mouse igg1 fc domain
    Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human <t>IgG</t> secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with the Fc region of mouse IgG1 at the C terminus
    https://www.bioz.com/result/mouse igg1 fc domain/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse igg1 fc domain - by Bioz Stars, 2021-09
    95/100 stars

    Images

    1) Product Images from "Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein"

    Article Title: Rapid isolation and profiling of a diverse panel of human monoclonal antibodies targeting the SARS-CoV-2 spike protein

    Journal: bioRxiv

    doi: 10.1101/2020.05.12.091462

    Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human IgG secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.
    Figure Legend Snippet: Functional assays from single antigen-reactive B cells. a. Schematic of detection of antigen-specific antibody . Biotinylated antigen (dark grey) was coupled to a streptavidin-conjugated polystyrene bead (light grey). Antibodies (blue) are secreted by single B cells loaded into individual NanoPens on the Berkeley Lights Beacon optofluidic device. Antibody binding to antigen was detected with a fluorescent anti-human IgG secondary Ab (black). b. Left : Schematic of fluorescing beads in the channel above a pen containing an individual B cell indicates antigen-specific reactivity. Top right : False-color still image of positive wells with B cells secreting S2P ecto -reactive antibodies. Reactive antibody diffusing out of a pen is visualized as a plume of fluorescence. Bottom right : False-color still image of positive wells with B cells secreting RBD-mFc-reactive antibodies. c . Representative images of RBD-mFc reactive clones.

    Techniques Used: Functional Assay, Binding Assay, Fluorescence, Clone Assay

    Related Articles

    Recombinant:

    Article Title: A SARS-CoV-2 neutralizing antibody with extensive Spike binding coverage and modified for optimal therapeutic outcomes
    Article Snippet: .. The 384-well plate (Corning, Catalog #3700) was coated overnight at 4 °C with PBS containing 30 μL 20 nM of the SARS-CoV-2 Spike S1+S2 ECD, his Tag protein (Sino Biological, Catalog #40589-V08B1), or SARS-CoV-2 Spike RBD-mFc recombinant protein (Sino Biological, Catalog #40592-V05H) or S1-mFc (Sino Biological, Catalog #40591-V05H1). ..

    Article Title: Identification of Natural Compounds as SARS-CoV-2 Entry Inhibitors by Molecular Docking-based Virtual Screening with Bio-layer Interferometry
    Article Snippet: .. 3 Recombinant proteinsMouse Fc-tagged RBD (Catalog: 40592-V05H) and His-tagged human ACE2 (Cat: 10108-H08B) were both purchased from Sino Biological (Beijing, China). ..

    Article Title: A SARS-CoV-2 neutralizing antibody with extensive Spike binding coverage and modified for optimal therapeutic outcomes
    Article Snippet: .. Convalescent COVID-19 patient antibody titers against SARS-CoV-2 S proteinsThe 384-well plates were coated overnight at 4 °C with PBS containing 1 µg/mL of SARS-CoV-2 Spike RBD-mFc recombinant protein (Sino Biological, Catalog #40592-V05H) or full-length S-his (Sino Biological, Catalog #40589-V08B1) or S1-mFc (Sino Biological, Catalog #40591-V05H1) or S1-his (Kactus, Catalog #COV-VM4S1). ..

    Article Title: Three epitope-distinct human antibodies from RenMab mice neutralize SARS-CoV-2 and cooperatively minimize the escape of mutants
    Article Snippet: .. In brief, 20 μg recombinant SARS-Cov-2 RBD (40592-V05H; Sino Biological) was emulsified in complete Freund’s adjuvant (CFA) and injected subcutaneously for the first immunization. ..

    Competitive ELISA:

    Article Title: Development and utilization of a surrogate SARS-CoV-2 viral neutralization assay to assess mRNA vaccine responses
    Article Snippet: .. Competitive ELISA Ninety six well Nunc Maxisorp flat-bottom plates were coated with 50 μL/well of murine gamma-immunoglobulin constant region (mFc) dimerized SARS-CoV-2 (2019-nCoV) Spike Protein RBD (mFc-RBD) from Sino Biological (Chesterbrook, PA), catalog number: 40592-V05H, at 2 μg/mL in 1 x phosphate buffered saline (PBS) at 4°C overnight. ..

    Selection:

    Article Title: A human antibody of potent efficacy against SARS-CoV-2 in rhesus macaques showed strong blocking activity to B.1.351
    Article Snippet: .. The phage selection was carried out in solution using 10 μg of biotinylated SARS-CoV-2 RBD protein (Sino Biological, 40592-V05H). ..

    Mouse Assay:

    Article Title: Structure and function analysis of a potent human neutralizing antibody CA521FALA against SARS-CoV-2
    Article Snippet: .. Another two mice (named Q14, Q15) were immunized in 10-day intervals with mixtures of Spike S1 protein (Cat 40591-V02H, Sino Biological) and RBD protein (Cat 40592-V05H, Sino Biological) and boosted by 70 μg mixtures (35 μg for each protein) after three rounds immunization. ..

    Incubation:

    Article Title: Discovery of Small Anti‐ACE2 Peptides to Inhibit SARS‐CoV‐2 Infectivity
    Article Snippet: .. The first well was blocked with BSA (2%) for 2 h, followed by incubation with 2019‐nCov Spike Protein RBD (500 ng) (catalog# 40592‐V05H, Sino Biological, Wayne, PA) for 2 h. The Ph.D.‐12 Phage Display Peptide Library (catalog # E81102, New England BioLabs, Ipswich, MA) was added to the first well and incubated at room temperature for 1 h. Unbound phages from the first well were then transferred to the second well and incubated for 1 h. The bound phages were eluted and amplified for the next round of biopanning. ..

    Amplification:

    Article Title: Discovery of Small Anti‐ACE2 Peptides to Inhibit SARS‐CoV‐2 Infectivity
    Article Snippet: .. The first well was blocked with BSA (2%) for 2 h, followed by incubation with 2019‐nCov Spike Protein RBD (500 ng) (catalog# 40592‐V05H, Sino Biological, Wayne, PA) for 2 h. The Ph.D.‐12 Phage Display Peptide Library (catalog # E81102, New England BioLabs, Ipswich, MA) was added to the first well and incubated at room temperature for 1 h. Unbound phages from the first well were then transferred to the second well and incubated for 1 h. The bound phages were eluted and amplified for the next round of biopanning. ..

    Injection:

    Article Title: Three epitope-distinct human antibodies from RenMab mice neutralize SARS-CoV-2 and cooperatively minimize the escape of mutants
    Article Snippet: .. In brief, 20 μg recombinant SARS-Cov-2 RBD (40592-V05H; Sino Biological) was emulsified in complete Freund’s adjuvant (CFA) and injected subcutaneously for the first immunization. ..

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  • 99
    Sino Biological sars cov 2 2019 ncov spike s1 s2 ecd his recombinant protein covid 19 spike research
    Adjuvanted S and S1 immune sera exhibit high titers of <t>SARS-CoV-2</t> pseudovirus neutralization and receptor binding inhibition activities. ( A – C ) Reduction percentage (%) in relative luminometer units (RLU) as a measure of luciferase activity for SARS-CoV-2 spike pseudotyped lentivirus infection in HEK293 cells expressing human ACE2 receptor. Data were obtained from pooled sera ( n = 6 to 8) with triplicate wells. S-0.8 (y): S 0.8 µg boost sera of young adult mice, S-0.8 + adj (y): S 0.8 µg + adjuvant boost sera of young adult mice, S-0.8 + adj (a): S 0.8 µg + adjuvant boost sera of old aged mice, S1-4 (y): S1 4 µg boost immune sera of young adult mice, S1-4 + adj (y): S1 4 µg + adjuvant boost immune sera of young adult mice, S2-4 (y): S2 4 µg boost immune sera of young adult mice, S2-4 + adj (y): S2 4 µg + adjuvant boost immune sera of young adult mice, S-0.8 + adj (y, x3): S 0.8 µg + adjuvant 2nd boost immune sera of young adult mice, S-0.8 + adj (y, x3, 19W): S 0.8 µg + adjuvant immune sera collected at week 19 post 2nd boost of young adult mice, S-4 + adj (a, x3): S 4 µg + adjuvant 2nd boost sera of old aged mice. IV-0.8-10 + adj (y, x3): inactivated adjuvanted SARS-CoV-2 vaccination in young age mice (prime 0.8 µg of heat-inactivated and gamma-irradiated virus, 2 times boost with 10 µg inactivated adjuvanted SARS-CoV-2 of heat-inactivated and gamma-irradiated virus). Adj: adjuvants (MPL + QS-21, 1 µg + 10 µg). Mock: sera from mice with adjuvant (MPL + QS-21, 1 + 10 µg) only. ( D – F ) ACE2 receptor binding inhibition titers in pooled immune sera ( n = 6–8) with triplicate wells. Inhibition percentage (%) of hACE2 binding to RBD was measured after incubation with serially diluted immune sera in the plate precoated with hACE2 protein. Immune sera of groups are the same as in ( A – C ). Statistical significance was calculated using two-way ANOVA and a Bonferroni’s multiple-comparison test. Error bars indicate the mean ± SEM. **; p
    Sars Cov 2 2019 Ncov Spike S1 S2 Ecd His Recombinant Protein Covid 19 Spike Research, supplied by Sino Biological, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 2019 ncov spike s1 s2 ecd his recombinant protein covid 19 spike research/product/Sino Biological
    Average 99 stars, based on 1 article reviews
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    86
    sino biological sars cov 2 spike protein
    Correlative analysis of <t>anti-SARS-CoV-2</t> immunoassays. Assays were performed to detect anti spike- Immunoglobulin (Ig) using the Ortho-Vitros assay (represented by signal to cut-off ratios (S/CO)), or anti-spike IgG with ELISA (represented by area under the curve, AUC), anti-RBD Ig using the Lumit Dx assay (represented by sample signal to calibrator ratios (S/C)) and 50% neutralizing titers with a retroviral-pseudotype assay (represented by IC50). Analysis of correlation were performed using Spearman’s correlation test for (A) anti-RBD titer, S/C vs IC50, (B) total anti-S Ig titer, S/CO vs IC50 and(C) anti-RBD titer, S/C vs total anti-S Ig titer, S/CO. These assay results were also qualified by comparing IC50 (D), anti-RBD titer, S/C (E) and total anti-S Ig titer, S/CO (F) against an in-house anti-spike protein IgG ELISA. Spearman’s correlation coefficient, ρ, and p values are indicated for each correlation.
    Sars Cov 2 Spike Protein, supplied by sino biological, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Sino Biological recombinant sars cov 2 s protein
    Ag-specific proliferation of <t>SARS-CoV-2</t> S-specific T cells. Proliferation assay using splenocytes of mice vaccinated on days 0 and 28 with indicated viruses, isolated 21 days after boost immunization, after co-culture with DC2.4 dendritic cell line transgenic for SARS-CoV-2 S (SARS2-S) or untransduced parental DC2.4 (untr.). Depicted are the percentages of ( A ) CD4 + or ( B ) CD8 + T cells with low CFSE staining, indicating proliferation in the samples. To analyze cellular α-MeV responses, splenocytes were stimulated with 10 μg/ml MeV bulk antigens or were left unstimulated (medium). The reactivity of splenocytes was confirmed by concanavalin A (ConA) treatment (10 μg/ml). Results for splenocytes of vaccinated mice are displayed individually and the trend between paired unstimulated and re-stimulated samples is outlined (n = 2-4). One-tailed Mann-Whitney t-test. *, p
    Recombinant Sars Cov 2 S Protein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant sars cov 2 s protein/product/Sino Biological
    Average 98 stars, based on 1 article reviews
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    Image Search Results


    Adjuvanted S and S1 immune sera exhibit high titers of SARS-CoV-2 pseudovirus neutralization and receptor binding inhibition activities. ( A – C ) Reduction percentage (%) in relative luminometer units (RLU) as a measure of luciferase activity for SARS-CoV-2 spike pseudotyped lentivirus infection in HEK293 cells expressing human ACE2 receptor. Data were obtained from pooled sera ( n = 6 to 8) with triplicate wells. S-0.8 (y): S 0.8 µg boost sera of young adult mice, S-0.8 + adj (y): S 0.8 µg + adjuvant boost sera of young adult mice, S-0.8 + adj (a): S 0.8 µg + adjuvant boost sera of old aged mice, S1-4 (y): S1 4 µg boost immune sera of young adult mice, S1-4 + adj (y): S1 4 µg + adjuvant boost immune sera of young adult mice, S2-4 (y): S2 4 µg boost immune sera of young adult mice, S2-4 + adj (y): S2 4 µg + adjuvant boost immune sera of young adult mice, S-0.8 + adj (y, x3): S 0.8 µg + adjuvant 2nd boost immune sera of young adult mice, S-0.8 + adj (y, x3, 19W): S 0.8 µg + adjuvant immune sera collected at week 19 post 2nd boost of young adult mice, S-4 + adj (a, x3): S 4 µg + adjuvant 2nd boost sera of old aged mice. IV-0.8-10 + adj (y, x3): inactivated adjuvanted SARS-CoV-2 vaccination in young age mice (prime 0.8 µg of heat-inactivated and gamma-irradiated virus, 2 times boost with 10 µg inactivated adjuvanted SARS-CoV-2 of heat-inactivated and gamma-irradiated virus). Adj: adjuvants (MPL + QS-21, 1 µg + 10 µg). Mock: sera from mice with adjuvant (MPL + QS-21, 1 + 10 µg) only. ( D – F ) ACE2 receptor binding inhibition titers in pooled immune sera ( n = 6–8) with triplicate wells. Inhibition percentage (%) of hACE2 binding to RBD was measured after incubation with serially diluted immune sera in the plate precoated with hACE2 protein. Immune sera of groups are the same as in ( A – C ). Statistical significance was calculated using two-way ANOVA and a Bonferroni’s multiple-comparison test. Error bars indicate the mean ± SEM. **; p

    Journal: Vaccines

    Article Title: Immunogenicity and Neutralizing Activity Comparison of SARS-CoV-2 Spike Full-Length and Subunit Domain Proteins in Young Adult and Old-Aged Mice

    doi: 10.3390/vaccines9040316

    Figure Lengend Snippet: Adjuvanted S and S1 immune sera exhibit high titers of SARS-CoV-2 pseudovirus neutralization and receptor binding inhibition activities. ( A – C ) Reduction percentage (%) in relative luminometer units (RLU) as a measure of luciferase activity for SARS-CoV-2 spike pseudotyped lentivirus infection in HEK293 cells expressing human ACE2 receptor. Data were obtained from pooled sera ( n = 6 to 8) with triplicate wells. S-0.8 (y): S 0.8 µg boost sera of young adult mice, S-0.8 + adj (y): S 0.8 µg + adjuvant boost sera of young adult mice, S-0.8 + adj (a): S 0.8 µg + adjuvant boost sera of old aged mice, S1-4 (y): S1 4 µg boost immune sera of young adult mice, S1-4 + adj (y): S1 4 µg + adjuvant boost immune sera of young adult mice, S2-4 (y): S2 4 µg boost immune sera of young adult mice, S2-4 + adj (y): S2 4 µg + adjuvant boost immune sera of young adult mice, S-0.8 + adj (y, x3): S 0.8 µg + adjuvant 2nd boost immune sera of young adult mice, S-0.8 + adj (y, x3, 19W): S 0.8 µg + adjuvant immune sera collected at week 19 post 2nd boost of young adult mice, S-4 + adj (a, x3): S 4 µg + adjuvant 2nd boost sera of old aged mice. IV-0.8-10 + adj (y, x3): inactivated adjuvanted SARS-CoV-2 vaccination in young age mice (prime 0.8 µg of heat-inactivated and gamma-irradiated virus, 2 times boost with 10 µg inactivated adjuvanted SARS-CoV-2 of heat-inactivated and gamma-irradiated virus). Adj: adjuvants (MPL + QS-21, 1 µg + 10 µg). Mock: sera from mice with adjuvant (MPL + QS-21, 1 + 10 µg) only. ( D – F ) ACE2 receptor binding inhibition titers in pooled immune sera ( n = 6–8) with triplicate wells. Inhibition percentage (%) of hACE2 binding to RBD was measured after incubation with serially diluted immune sera in the plate precoated with hACE2 protein. Immune sera of groups are the same as in ( A – C ). Statistical significance was calculated using two-way ANOVA and a Bonferroni’s multiple-comparison test. Error bars indicate the mean ± SEM. **; p

    Article Snippet: Recombinant Proteins and ReagentsSARS-CoV-2 different recombinant S and receptor proteins were obtained from Sino Biologicals (Wayne, PA, USA): Full-length S (S1–S2) ectodomain amino acid (aa) residues 16-1213 (40589-V08B1, 134.36 kDa, expressed in baculovirus-insect cells), S1 subunit (aa 16-685) with RBD domain (40591-V08H, 76.5 kDa, expressed in HEK293 cells); S2 subunit (aa 686-1213) with fusion domain (40589-V08B1, 59.36 kDa, expressed in baculovirus-insect cells); human angiotensin-converting enzyme 2 (hACE2) protein (aa 1-740) fused to Fc tag (10108-H02H, expressed in HEK293 cells).

    Techniques: Neutralization, Binding Assay, Inhibition, Luciferase, Activity Assay, Infection, Expressing, Mouse Assay, Irradiation, Incubation

    B cell and T cell immune responses to SARS-CoV-2 S vaccination in young adult and old aged mice. To determine cellular immunity, spleen cells were prepared from immunized young adult ( n = 6) and old aged mice ( n = 8). ( A ) Antibody-secreting cells (ASCs) specific for full-length S protein were determined on the ELISpot plate precoated with full-length S protein. ( B ) IFN-γ-secreting cells were analyzed by in vitro stimulation with pooled S peptides or full-length S protein using ELISpot assay. ( C , D ). IFN-γ + CD4 and IFN-γ + CD8 T cells were determined by flow cytometry after in vitro stimulation with pooled S peptides and intracellular cytokine antibi staining. S-0.8 (y): S 0.8 µg vaccination of young adult mice, S-0.8 + adj (y): S 0.8 µg + adjuvant vaccination of young adult mice, S-0.8 + adj (a): S 0.8 µg + adjuvant vaccination of old aged mice. Mock: sera from mice with adjuvant (MPL + QS-21, 1 + 10 µg) only. Statistical significance was calculated using one-way ANOVA and a Dunnett’s multiple-comparison test. Error bars indicate the mean ± SEM. *; p

    Journal: Vaccines

    Article Title: Immunogenicity and Neutralizing Activity Comparison of SARS-CoV-2 Spike Full-Length and Subunit Domain Proteins in Young Adult and Old-Aged Mice

    doi: 10.3390/vaccines9040316

    Figure Lengend Snippet: B cell and T cell immune responses to SARS-CoV-2 S vaccination in young adult and old aged mice. To determine cellular immunity, spleen cells were prepared from immunized young adult ( n = 6) and old aged mice ( n = 8). ( A ) Antibody-secreting cells (ASCs) specific for full-length S protein were determined on the ELISpot plate precoated with full-length S protein. ( B ) IFN-γ-secreting cells were analyzed by in vitro stimulation with pooled S peptides or full-length S protein using ELISpot assay. ( C , D ). IFN-γ + CD4 and IFN-γ + CD8 T cells were determined by flow cytometry after in vitro stimulation with pooled S peptides and intracellular cytokine antibi staining. S-0.8 (y): S 0.8 µg vaccination of young adult mice, S-0.8 + adj (y): S 0.8 µg + adjuvant vaccination of young adult mice, S-0.8 + adj (a): S 0.8 µg + adjuvant vaccination of old aged mice. Mock: sera from mice with adjuvant (MPL + QS-21, 1 + 10 µg) only. Statistical significance was calculated using one-way ANOVA and a Dunnett’s multiple-comparison test. Error bars indicate the mean ± SEM. *; p

    Article Snippet: Recombinant Proteins and ReagentsSARS-CoV-2 different recombinant S and receptor proteins were obtained from Sino Biologicals (Wayne, PA, USA): Full-length S (S1–S2) ectodomain amino acid (aa) residues 16-1213 (40589-V08B1, 134.36 kDa, expressed in baculovirus-insect cells), S1 subunit (aa 16-685) with RBD domain (40591-V08H, 76.5 kDa, expressed in HEK293 cells); S2 subunit (aa 686-1213) with fusion domain (40589-V08B1, 59.36 kDa, expressed in baculovirus-insect cells); human angiotensin-converting enzyme 2 (hACE2) protein (aa 1-740) fused to Fc tag (10108-H02H, expressed in HEK293 cells).

    Techniques: Mouse Assay, Enzyme-linked Immunospot, In Vitro, Flow Cytometry, Staining

    SARS-CoV-2 full-length spike (S) ectodomain and subunit proteins and receptor binding activities. ( A ) Full-length S (S1–S2) ectodomain contains aa residues 16-1213, S1 subunit aa 16-685 (green), and S2 subunit aa 686-1213. NTD: N-terminal domain (blue), RBD: receptor binding domain. ( B , C ) The receptor binding properties were determined using serially diluted soluble hACE2-Fc (0.5–2 µg/mL) on the 96-well plates precoated with 0.8 µg ( B ) or 2 µg ( C ) of S (S1–S2) and S1 subunit proteins. Due to different molecular masses of S and S1 proteins despite the same concentration, molarity in nanomoles (nM) is indicated for each protein coated.

    Journal: Vaccines

    Article Title: Immunogenicity and Neutralizing Activity Comparison of SARS-CoV-2 Spike Full-Length and Subunit Domain Proteins in Young Adult and Old-Aged Mice

    doi: 10.3390/vaccines9040316

    Figure Lengend Snippet: SARS-CoV-2 full-length spike (S) ectodomain and subunit proteins and receptor binding activities. ( A ) Full-length S (S1–S2) ectodomain contains aa residues 16-1213, S1 subunit aa 16-685 (green), and S2 subunit aa 686-1213. NTD: N-terminal domain (blue), RBD: receptor binding domain. ( B , C ) The receptor binding properties were determined using serially diluted soluble hACE2-Fc (0.5–2 µg/mL) on the 96-well plates precoated with 0.8 µg ( B ) or 2 µg ( C ) of S (S1–S2) and S1 subunit proteins. Due to different molecular masses of S and S1 proteins despite the same concentration, molarity in nanomoles (nM) is indicated for each protein coated.

    Article Snippet: Recombinant Proteins and ReagentsSARS-CoV-2 different recombinant S and receptor proteins were obtained from Sino Biologicals (Wayne, PA, USA): Full-length S (S1–S2) ectodomain amino acid (aa) residues 16-1213 (40589-V08B1, 134.36 kDa, expressed in baculovirus-insect cells), S1 subunit (aa 16-685) with RBD domain (40591-V08H, 76.5 kDa, expressed in HEK293 cells); S2 subunit (aa 686-1213) with fusion domain (40589-V08B1, 59.36 kDa, expressed in baculovirus-insect cells); human angiotensin-converting enzyme 2 (hACE2) protein (aa 1-740) fused to Fc tag (10108-H02H, expressed in HEK293 cells).

    Techniques: Binding Assay, Concentration Assay

    Correlative analysis of anti-SARS-CoV-2 immunoassays. Assays were performed to detect anti spike- Immunoglobulin (Ig) using the Ortho-Vitros assay (represented by signal to cut-off ratios (S/CO)), or anti-spike IgG with ELISA (represented by area under the curve, AUC), anti-RBD Ig using the Lumit Dx assay (represented by sample signal to calibrator ratios (S/C)) and 50% neutralizing titers with a retroviral-pseudotype assay (represented by IC50). Analysis of correlation were performed using Spearman’s correlation test for (A) anti-RBD titer, S/C vs IC50, (B) total anti-S Ig titer, S/CO vs IC50 and(C) anti-RBD titer, S/C vs total anti-S Ig titer, S/CO. These assay results were also qualified by comparing IC50 (D), anti-RBD titer, S/C (E) and total anti-S Ig titer, S/CO (F) against an in-house anti-spike protein IgG ELISA. Spearman’s correlation coefficient, ρ, and p values are indicated for each correlation.

    Journal: PLoS ONE

    Article Title: Predicting the efficacy of COVID-19 convalescent plasma donor units with the Lumit Dx anti-receptor binding domain assay

    doi: 10.1371/journal.pone.0253551

    Figure Lengend Snippet: Correlative analysis of anti-SARS-CoV-2 immunoassays. Assays were performed to detect anti spike- Immunoglobulin (Ig) using the Ortho-Vitros assay (represented by signal to cut-off ratios (S/CO)), or anti-spike IgG with ELISA (represented by area under the curve, AUC), anti-RBD Ig using the Lumit Dx assay (represented by sample signal to calibrator ratios (S/C)) and 50% neutralizing titers with a retroviral-pseudotype assay (represented by IC50). Analysis of correlation were performed using Spearman’s correlation test for (A) anti-RBD titer, S/C vs IC50, (B) total anti-S Ig titer, S/CO vs IC50 and(C) anti-RBD titer, S/C vs total anti-S Ig titer, S/CO. These assay results were also qualified by comparing IC50 (D), anti-RBD titer, S/C (E) and total anti-S Ig titer, S/CO (F) against an in-house anti-spike protein IgG ELISA. Spearman’s correlation coefficient, ρ, and p values are indicated for each correlation.

    Article Snippet: The Promega Lumit Dx SARS-CoV-2 Immunoassay was used according to the manufacturer’s protocol for detection of antibody binding to the RBD of SARS-CoV-2 spike protein.

    Techniques: Enzyme-linked Immunosorbent Assay, CytoScan DX Assay

    Ag-specific proliferation of SARS-CoV-2 S-specific T cells. Proliferation assay using splenocytes of mice vaccinated on days 0 and 28 with indicated viruses, isolated 21 days after boost immunization, after co-culture with DC2.4 dendritic cell line transgenic for SARS-CoV-2 S (SARS2-S) or untransduced parental DC2.4 (untr.). Depicted are the percentages of ( A ) CD4 + or ( B ) CD8 + T cells with low CFSE staining, indicating proliferation in the samples. To analyze cellular α-MeV responses, splenocytes were stimulated with 10 μg/ml MeV bulk antigens or were left unstimulated (medium). The reactivity of splenocytes was confirmed by concanavalin A (ConA) treatment (10 μg/ml). Results for splenocytes of vaccinated mice are displayed individually and the trend between paired unstimulated and re-stimulated samples is outlined (n = 2-4). One-tailed Mann-Whitney t-test. *, p

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Ag-specific proliferation of SARS-CoV-2 S-specific T cells. Proliferation assay using splenocytes of mice vaccinated on days 0 and 28 with indicated viruses, isolated 21 days after boost immunization, after co-culture with DC2.4 dendritic cell line transgenic for SARS-CoV-2 S (SARS2-S) or untransduced parental DC2.4 (untr.). Depicted are the percentages of ( A ) CD4 + or ( B ) CD8 + T cells with low CFSE staining, indicating proliferation in the samples. To analyze cellular α-MeV responses, splenocytes were stimulated with 10 μg/ml MeV bulk antigens or were left unstimulated (medium). The reactivity of splenocytes was confirmed by concanavalin A (ConA) treatment (10 μg/ml). Results for splenocytes of vaccinated mice are displayed individually and the trend between paired unstimulated and re-stimulated samples is outlined (n = 2-4). One-tailed Mann-Whitney t-test. *, p

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: Proliferation Assay, Mouse Assay, Isolation, Co-Culture Assay, Transgenic Assay, Staining, One-tailed Test, MANN-WHITNEY

    Antigen-specific killing activity of SARS-CoV-2 S-specific T cells. Killing assay using splenocytes of mice vaccinated on days 0 and 28 isolated 21 days after the second immunization. Splenocytes were co-cultured with DC2.4 ( A ) or with antigen-presenting DC2.4-SARS2-S ( B ) cells or for 6 days. Activated CTLs were then co-cultured with EL-4 green -SARS2-S target cells (Antigen) and EL-4 red control cells (NC) at indicated E:T ratios for 4 h. Ratio of living target to non-target cells (Antigen:NC) was determined by flow cytometry. Depicted are means and standard deviation of each group (open diamonds, MeV vac2 -SARS2-S(H); filled circles, mock; filled squares, MV vac2 -ATU(P); grey triangles: S protein + Alum) (n = 3 - 5). For statistical analysis of grouped ELISpot data, two-way ANOVA analysis was applied with paired Tukey’s Multi comparison test used as post hoc test. ****, p

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Antigen-specific killing activity of SARS-CoV-2 S-specific T cells. Killing assay using splenocytes of mice vaccinated on days 0 and 28 isolated 21 days after the second immunization. Splenocytes were co-cultured with DC2.4 ( A ) or with antigen-presenting DC2.4-SARS2-S ( B ) cells or for 6 days. Activated CTLs were then co-cultured with EL-4 green -SARS2-S target cells (Antigen) and EL-4 red control cells (NC) at indicated E:T ratios for 4 h. Ratio of living target to non-target cells (Antigen:NC) was determined by flow cytometry. Depicted are means and standard deviation of each group (open diamonds, MeV vac2 -SARS2-S(H); filled circles, mock; filled squares, MV vac2 -ATU(P); grey triangles: S protein + Alum) (n = 3 - 5). For statistical analysis of grouped ELISpot data, two-way ANOVA analysis was applied with paired Tukey’s Multi comparison test used as post hoc test. ****, p

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: Activity Assay, Mouse Assay, Isolation, Cell Culture, Flow Cytometry, Standard Deviation, Enzyme-linked Immunospot

    Induction of a-SARS-CoV-2 S and a-MeV specific antibodies. Sera of mice vaccinated on days 0 and 28 with indicated viruses or Alum-adjuvanted S protein were sampled on day 0 (A, D, E, F), day 28 after prime- (B, E, H, K) and day 49 after boost-immunization (C, F, I, L) and analyzed for antibodies specific for SARS-CoV-2 S or MeV. Medium-inoculated mice served as mock. Pan-IgG binding to recombinant SARS-CoV S (A – C) or MeV bulk antigens (G – I) were determined by ELISA via the specific OD 450 nm value. Depicted are means and respective standard deviation of the mean (SEM) of each group (n = 5 - 6). Virus neutralizing titers (VNT) in vaccinated mice for SARS-CoV-2 (D - F) or MeV (J – L) were calculated as reciprocal of the highest dilution abolishing infectivity. ( M) SARS-CoV-2 VNT of 4 human Covid-19 reconvalescent sera. Dots represent single individuals; horizontal line represents mean per group. For statistical analysis of VNT data, one-way ANOVA was performed in combination with Tukey’s Multi comparison test to compare all pair means.

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Induction of a-SARS-CoV-2 S and a-MeV specific antibodies. Sera of mice vaccinated on days 0 and 28 with indicated viruses or Alum-adjuvanted S protein were sampled on day 0 (A, D, E, F), day 28 after prime- (B, E, H, K) and day 49 after boost-immunization (C, F, I, L) and analyzed for antibodies specific for SARS-CoV-2 S or MeV. Medium-inoculated mice served as mock. Pan-IgG binding to recombinant SARS-CoV S (A – C) or MeV bulk antigens (G – I) were determined by ELISA via the specific OD 450 nm value. Depicted are means and respective standard deviation of the mean (SEM) of each group (n = 5 - 6). Virus neutralizing titers (VNT) in vaccinated mice for SARS-CoV-2 (D - F) or MeV (J – L) were calculated as reciprocal of the highest dilution abolishing infectivity. ( M) SARS-CoV-2 VNT of 4 human Covid-19 reconvalescent sera. Dots represent single individuals; horizontal line represents mean per group. For statistical analysis of VNT data, one-way ANOVA was performed in combination with Tukey’s Multi comparison test to compare all pair means.

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: Mouse Assay, Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Standard Deviation, Infection

    Immune bias of induced responses. To analyze skewing of immune responses towards Th1- or Th2-biased immunity ( A ) sera and ( B ) splenocytes of vaccinated mice depicted before were analyzed. ( A ) Sera of mice vaccinated on days 0 and 28 with MeV vac2 -SARS2-S(H) or Alum-adjuvanted S protein already shown in Fig. 2 were analysed for IgG1- or IgG2a-type antibodies specific for SARS-CoV-2 S. IgG1 (left panel) or IgG2a (right panel) binding to recombinant SARS-CoV S were determined by ELISA via the specific OD 450 nm value. Depicted are means and respective standard deviation of the mean (SEM) of each group (n = 5 - 6). ( B ) Splenocytes of the same mice already shown in Figs. 3 to 6 were analysed by multiplex cytokine analysis for secretion of typical marker cytokines in the supernatant after re-stimulation by co-culture with antigen-presenting DC2.4-SARS2-S cells. DC2.4 cells served as non-specific control stimulus. Dots represent individual animals, horizontal bars mean per group (n = 4 - 5). IFN-γ: upper limit of detection (ULOD): 2015.2 pg/mL; IL-6: ULOD: 3992,4 pg/mL; IL-17a lower limit of detection (LLOD): 0.473 pg/mL; IL-4 LLOD: 0.095 pg/mL; IL-5 LLOD: 0.685 pg/mL; IL-13 LLOD: 3.463 pg/mL. For statistical analysis of grouped multiplex data, two-way ANOVA analysis was applied with paired Tukey’s Multi comparison test as post hoc test. *, p

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Immune bias of induced responses. To analyze skewing of immune responses towards Th1- or Th2-biased immunity ( A ) sera and ( B ) splenocytes of vaccinated mice depicted before were analyzed. ( A ) Sera of mice vaccinated on days 0 and 28 with MeV vac2 -SARS2-S(H) or Alum-adjuvanted S protein already shown in Fig. 2 were analysed for IgG1- or IgG2a-type antibodies specific for SARS-CoV-2 S. IgG1 (left panel) or IgG2a (right panel) binding to recombinant SARS-CoV S were determined by ELISA via the specific OD 450 nm value. Depicted are means and respective standard deviation of the mean (SEM) of each group (n = 5 - 6). ( B ) Splenocytes of the same mice already shown in Figs. 3 to 6 were analysed by multiplex cytokine analysis for secretion of typical marker cytokines in the supernatant after re-stimulation by co-culture with antigen-presenting DC2.4-SARS2-S cells. DC2.4 cells served as non-specific control stimulus. Dots represent individual animals, horizontal bars mean per group (n = 4 - 5). IFN-γ: upper limit of detection (ULOD): 2015.2 pg/mL; IL-6: ULOD: 3992,4 pg/mL; IL-17a lower limit of detection (LLOD): 0.473 pg/mL; IL-4 LLOD: 0.095 pg/mL; IL-5 LLOD: 0.685 pg/mL; IL-13 LLOD: 3.463 pg/mL. For statistical analysis of grouped multiplex data, two-way ANOVA analysis was applied with paired Tukey’s Multi comparison test as post hoc test. *, p

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: Mouse Assay, Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Standard Deviation, Multiplex Assay, Marker, Co-Culture Assay

    Characterization of fusogenic phenotype of MeV vac2 -SARS2-S(P) and MeV vac2 -SARS2-S(H). (A ) Photographs of fusion activity of Vero cells infected at an MOI of 0.01 with MeV vac2 -SARS2-S(P) or MeVvac2-SARS2-S(H) encoding SARS-CoV-2 S in additional transcription units post P or post H, respectively, in direct comparison to MV vac2 -ATU(P) or MV vac2 -GFP(H) control vaccine viruses or MV NSe -GFP(N) hyperfusogenic oncolytic MeV. Representative picture of one out of three independent experiments. Scale bar represents 200 mm. ( B ) Cell fusion was quantified 30 h after infection. For statistical analysis, one-way ANOVA was performed in combination with Tukey’s Multi comparison test to compare all pair means. *, p

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Characterization of fusogenic phenotype of MeV vac2 -SARS2-S(P) and MeV vac2 -SARS2-S(H). (A ) Photographs of fusion activity of Vero cells infected at an MOI of 0.01 with MeV vac2 -SARS2-S(P) or MeVvac2-SARS2-S(H) encoding SARS-CoV-2 S in additional transcription units post P or post H, respectively, in direct comparison to MV vac2 -ATU(P) or MV vac2 -GFP(H) control vaccine viruses or MV NSe -GFP(N) hyperfusogenic oncolytic MeV. Representative picture of one out of three independent experiments. Scale bar represents 200 mm. ( B ) Cell fusion was quantified 30 h after infection. For statistical analysis, one-way ANOVA was performed in combination with Tukey’s Multi comparison test to compare all pair means. *, p

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: Activity Assay, Infection

    Generation and in vitro characterization of MeV vac2 -SARS2-S(P) and MeV vac2 -SARS2-S(H). (A) Schematic depiction of full-length SARS-CoV-2 S and recombinant MeV vac2 genomes used for expression of this antigen (lower schemes). Antigen or antigen encoding genes are depicted in dark grey; MeV viral gene cassettes (in light grey) are annotated. MluI and AatII restriction sites used for cloning of antigen-genes into post P or post H ATU are highlighted (B) Immunoblot analysis of Vero cells infected at an MOI of 0.01 with MeV vac2 -SARS2-S(P), MeV vac2 -SARS2-S(H), or MV vac2 -ATU(P) (MV vac2 ) as depicted above lanes. Uninfected cells served as mock. Blots were probed using rabbit polyclonal anti-SARS spike antibody (upper blot) or mAb reactive against MeV-N (lower blot). Arrows indicate specific bands. ( C, D ) Growth kinetics of recombinant MeV on Vero cells infected at an MOI of 0.03 with MV vac2 -ATU(P) or MeV vac2 -SARS2-S encoding extra genes in post H or post P. Titers of samples prepared at indicated time points post infection were titrated on Vero cells. Means and standard deviations of three to five independent experiments are presented. ( E ) SARS-CoV-2 S protein expression in Vero cells was verified via immunoperoxidase monolayer assay. 50× magnification; scale bar, 500 μm.

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Generation and in vitro characterization of MeV vac2 -SARS2-S(P) and MeV vac2 -SARS2-S(H). (A) Schematic depiction of full-length SARS-CoV-2 S and recombinant MeV vac2 genomes used for expression of this antigen (lower schemes). Antigen or antigen encoding genes are depicted in dark grey; MeV viral gene cassettes (in light grey) are annotated. MluI and AatII restriction sites used for cloning of antigen-genes into post P or post H ATU are highlighted (B) Immunoblot analysis of Vero cells infected at an MOI of 0.01 with MeV vac2 -SARS2-S(P), MeV vac2 -SARS2-S(H), or MV vac2 -ATU(P) (MV vac2 ) as depicted above lanes. Uninfected cells served as mock. Blots were probed using rabbit polyclonal anti-SARS spike antibody (upper blot) or mAb reactive against MeV-N (lower blot). Arrows indicate specific bands. ( C, D ) Growth kinetics of recombinant MeV on Vero cells infected at an MOI of 0.03 with MV vac2 -ATU(P) or MeV vac2 -SARS2-S encoding extra genes in post H or post P. Titers of samples prepared at indicated time points post infection were titrated on Vero cells. Means and standard deviations of three to five independent experiments are presented. ( E ) SARS-CoV-2 S protein expression in Vero cells was verified via immunoperoxidase monolayer assay. 50× magnification; scale bar, 500 μm.

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: In Vitro, Recombinant, Expressing, Clone Assay, Infection

    Expression of SARS-CoV-2 S protein in Vero and 293T cells. Photographic depiction of fusion activity in Vero or 293T cells 48 h after transfection with 1 µg of SARS-CoV-2 S expression plasmid of control DNA. One representative out of three independent experiments is shown. Scale bar represents 100 mm.

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Expression of SARS-CoV-2 S protein in Vero and 293T cells. Photographic depiction of fusion activity in Vero or 293T cells 48 h after transfection with 1 µg of SARS-CoV-2 S expression plasmid of control DNA. One representative out of three independent experiments is shown. Scale bar represents 100 mm.

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: Expressing, Activity Assay, Transfection, Plasmid Preparation

    Secretion of IFN-γ after antigen-specific re-stimulation of splenocytes. IFN-γ ELISpot analysis using splenocytes of mice vaccinated on days 0 and 28 with indicated vaccines, isolated 21 days after boost immunization, and after co-culture with DC2.4 or JAWSII dendritic cell lines transgenic for SARS-CoV-2 S (SARS2-S) or untransduced controls (untr.). To analyze cellular responses directed against MeV, splenocytes were stimulated with 10 μg/mL MeV bulk antigens or were left unstimulated as controls (medium). The reactivity of splenocytes was confirmed by Concanavalin A (ConA) treatment (10 μg/mL). The number of cells per 1×10 6 splenocytes represent the amount of cells expressing IFN-γ upon re-stimulation. Dots represent individual animals, horizontal bars mean per group (n = 5 - 6). Samples above the upper detection limit (ULOD) were displayed as such. For statistical analysis of grouped ELISpot data, two-way ANOVA analysis was applied with paired Tukey’s Multi comparison test used as post hoc test. ns, not significant (p > 0.05); ****, p

    Journal: bioRxiv

    Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

    doi: 10.1101/2020.07.11.198291

    Figure Lengend Snippet: Secretion of IFN-γ after antigen-specific re-stimulation of splenocytes. IFN-γ ELISpot analysis using splenocytes of mice vaccinated on days 0 and 28 with indicated vaccines, isolated 21 days after boost immunization, and after co-culture with DC2.4 or JAWSII dendritic cell lines transgenic for SARS-CoV-2 S (SARS2-S) or untransduced controls (untr.). To analyze cellular responses directed against MeV, splenocytes were stimulated with 10 μg/mL MeV bulk antigens or were left unstimulated as controls (medium). The reactivity of splenocytes was confirmed by Concanavalin A (ConA) treatment (10 μg/mL). The number of cells per 1×10 6 splenocytes represent the amount of cells expressing IFN-γ upon re-stimulation. Dots represent individual animals, horizontal bars mean per group (n = 5 - 6). Samples above the upper detection limit (ULOD) were displayed as such. For statistical analysis of grouped ELISpot data, two-way ANOVA analysis was applied with paired Tukey’s Multi comparison test used as post hoc test. ns, not significant (p > 0.05); ****, p

    Article Snippet: These animals were inoculated intraperitoneally (i.p.) with 1×105 TCID50 of recombinant vaccine viruses in 200 μl volume, or subcutaneously (s.c.) with 10 μg recombinant SARS-CoV-2 S protein (Sino Biological Europe, Eschborn, Germany) adjuvanted with 500 μg aluminium hydroxide (Alhydrogel adjuvant 2%, vac-alu-250, InvivoGen, San Diego, CA, USA) in 100 μl volume on days 0 and 28.

    Techniques: Enzyme-linked Immunospot, Mouse Assay, Isolation, Co-Culture Assay, Transgenic Assay, Expressing