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    Sino Biological sars cov 2 2019 ncov spike s1 his recombinant protein hplc verified covid 19 spike s1 research
    Sars Cov 2 2019 Ncov Spike S1 His Recombinant Protein Hplc Verified Covid 19 Spike S1 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 his recombinant protein hplc verified covid 19 spike s1 research/product/Sino Biological
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    86
    Sino Biological sars cov 2 2019 ncov spike rbd his recombinant protein covid 19 spike rbd research
    Concentration-dependent inhibition of <t>SARS-CoV-S1S2</t> binding to ACE2 by representative compounds of the present study. Concentration–response curves obtained for the inhibition of the PPI between SARS-CoV-S1S2 (His-tagged, 1 μg/mL) and hACE2 (Fc-conjugated, 1 μg/mL) in cell-free ELISA-type assay by selected representative dye and nondye compounds. Data and fit as before ( Figure 3 ). Most compounds including several DRI-C compounds show similar activity against SARS-CoV (i.e., SARS-CoV-1) as against <t>SARS-CoV-2</t> raising the possibility of broad-spectrum activity.
    Sars Cov 2 2019 Ncov Spike Rbd His Recombinant Protein Covid 19 Spike Rbd Research, 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
    https://www.bioz.com/result/sars cov 2 2019 ncov spike rbd his recombinant protein covid 19 spike rbd research/product/Sino Biological
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike rbd his recombinant protein covid 19 spike rbd research - by Bioz Stars, 2021-06
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    99
    Sino Biological sars cov 2 s protein
    Concentration-dependent inhibition of <t>SARS-CoV-S1S2</t> binding to ACE2 by representative compounds of the present study. Concentration–response curves obtained for the inhibition of the PPI between SARS-CoV-S1S2 (His-tagged, 1 μg/mL) and hACE2 (Fc-conjugated, 1 μg/mL) in cell-free ELISA-type assay by selected representative dye and nondye compounds. Data and fit as before ( Figure 3 ). Most compounds including several DRI-C compounds show similar activity against SARS-CoV (i.e., SARS-CoV-1) as against <t>SARS-CoV-2</t> raising the possibility of broad-spectrum activity.
    Sars Cov 2 S Protein, 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 s protein/product/Sino Biological
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 s protein - by Bioz Stars, 2021-06
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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
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike s1 s2 ecd his recombinant protein covid 19 spike research - by Bioz Stars, 2021-06
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    Image Search Results


    Concentration-dependent inhibition of SARS-CoV-S1S2 binding to ACE2 by representative compounds of the present study. Concentration–response curves obtained for the inhibition of the PPI between SARS-CoV-S1S2 (His-tagged, 1 μg/mL) and hACE2 (Fc-conjugated, 1 μg/mL) in cell-free ELISA-type assay by selected representative dye and nondye compounds. Data and fit as before ( Figure 3 ). Most compounds including several DRI-C compounds show similar activity against SARS-CoV (i.e., SARS-CoV-1) as against SARS-CoV-2 raising the possibility of broad-spectrum activity.

    Journal: ACS Infectious Diseases

    Article Title: Small-Molecule Inhibitors of the Coronavirus Spike: ACE2 Protein–Protein Interaction as Blockers of Viral Attachment and Entry for SARS-CoV-2

    doi: 10.1021/acsinfecdis.1c00070

    Figure Lengend Snippet: Concentration-dependent inhibition of SARS-CoV-S1S2 binding to ACE2 by representative compounds of the present study. Concentration–response curves obtained for the inhibition of the PPI between SARS-CoV-S1S2 (His-tagged, 1 μg/mL) and hACE2 (Fc-conjugated, 1 μg/mL) in cell-free ELISA-type assay by selected representative dye and nondye compounds. Data and fit as before ( Figure 3 ). Most compounds including several DRI-C compounds show similar activity against SARS-CoV (i.e., SARS-CoV-1) as against SARS-CoV-2 raising the possibility of broad-spectrum activity.

    Article Snippet: Binding Assays SARS-CoV-2 S1 and RBD (cat. no. 40591-V08H and 40592-V08H), SARS-CoV S1+S2 (cat. no. 40634-V08B), HCoV-NL63 S1 (cat. no. 40600-V08H; all with His tag), and ACE2-Fc (cat. no. 10108-H05H) used in the binding assay were obtained from SinoBiological (Wayne, PA, USA).

    Techniques: Concentration Assay, Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay

    Concentration–response curves for binding of CoV spike protein domains to human ACE2 in cell-free ELISA-type assays. Binding curves and corresponding EC 50 ’s are shown for SARS-CoV-2 (RBD and S1), SARS-CoV (S1 S2), and HCoV-NL63 (S1). They were obtained using Fc-conjugated hACE2 coated on the plate and His-tagged S1, S1S2, or RBD added in increasing amounts as shown with the amount bound detected using an anti-His–HRP conjugate (mean ± SD for two experiments in duplicates).

    Journal: ACS Infectious Diseases

    Article Title: Small-Molecule Inhibitors of the Coronavirus Spike: ACE2 Protein–Protein Interaction as Blockers of Viral Attachment and Entry for SARS-CoV-2

    doi: 10.1021/acsinfecdis.1c00070

    Figure Lengend Snippet: Concentration–response curves for binding of CoV spike protein domains to human ACE2 in cell-free ELISA-type assays. Binding curves and corresponding EC 50 ’s are shown for SARS-CoV-2 (RBD and S1), SARS-CoV (S1 S2), and HCoV-NL63 (S1). They were obtained using Fc-conjugated hACE2 coated on the plate and His-tagged S1, S1S2, or RBD added in increasing amounts as shown with the amount bound detected using an anti-His–HRP conjugate (mean ± SD for two experiments in duplicates).

    Article Snippet: Binding Assays SARS-CoV-2 S1 and RBD (cat. no. 40591-V08H and 40592-V08H), SARS-CoV S1+S2 (cat. no. 40634-V08B), HCoV-NL63 S1 (cat. no. 40600-V08H; all with His tag), and ACE2-Fc (cat. no. 10108-H05H) used in the binding assay were obtained from SinoBiological (Wayne, PA, USA).

    Techniques: Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay

    Concentration-dependent inhibition of SARS-CoV-2 pseudovirus entry (BacMam) into hACE2 expressing host cells by selected compounds. Quantification of entry of pseudoviruses bearing the SARS-CoV-2 S protein (plus green fluorescent protein reporters; BacMam-based) in ACE2 (plus red fluorescence)-expressing host cells (HEK293T). Representative images (bottom row) and their quantification for pseudovirus (green) and ACE2 expression (red) using ImageJ (top row) are shown from one experiment for CgRd and DRI-C23041 in (A) and (B), respectively; average data from three experiments fitted with typical concentration–response curves are shown in (C). The amount of green present is proportional with the number of infected cells as green fluorescence is expressed only in pseudovirus infected cells, while amount of red is proportional with the number of ACE2-expressing cells. The organic dye CgRd (A), but especially DRI-C23041 (B) showed concentration-dependent inhibition with activities corresponding to low micromolar IC 50 values, whereas hydroxychloroquine (HCQ) showed no effect (C).

    Journal: ACS Infectious Diseases

    Article Title: Small-Molecule Inhibitors of the Coronavirus Spike: ACE2 Protein–Protein Interaction as Blockers of Viral Attachment and Entry for SARS-CoV-2

    doi: 10.1021/acsinfecdis.1c00070

    Figure Lengend Snippet: Concentration-dependent inhibition of SARS-CoV-2 pseudovirus entry (BacMam) into hACE2 expressing host cells by selected compounds. Quantification of entry of pseudoviruses bearing the SARS-CoV-2 S protein (plus green fluorescent protein reporters; BacMam-based) in ACE2 (plus red fluorescence)-expressing host cells (HEK293T). Representative images (bottom row) and their quantification for pseudovirus (green) and ACE2 expression (red) using ImageJ (top row) are shown from one experiment for CgRd and DRI-C23041 in (A) and (B), respectively; average data from three experiments fitted with typical concentration–response curves are shown in (C). The amount of green present is proportional with the number of infected cells as green fluorescence is expressed only in pseudovirus infected cells, while amount of red is proportional with the number of ACE2-expressing cells. The organic dye CgRd (A), but especially DRI-C23041 (B) showed concentration-dependent inhibition with activities corresponding to low micromolar IC 50 values, whereas hydroxychloroquine (HCQ) showed no effect (C).

    Article Snippet: Binding Assays SARS-CoV-2 S1 and RBD (cat. no. 40591-V08H and 40592-V08H), SARS-CoV S1+S2 (cat. no. 40634-V08B), HCoV-NL63 S1 (cat. no. 40600-V08H; all with His tag), and ACE2-Fc (cat. no. 10108-H05H) used in the binding assay were obtained from SinoBiological (Wayne, PA, USA).

    Techniques: Concentration Assay, Inhibition, Expressing, Fluorescence, Infection

    Concentration-dependent inhibition of SARS-CoV-2 pseudovirus (VSV-Δ G ) entry into hACE2/Furin expressing host cells by selected compounds. Entry of VSV-ΔG pseudoviruses bearing the SARS-CoV-2 S protein (plus GFP reporters) in ACE2/Furin overexpressing host cells (Vero-E6) was quantified via GFP fluorescence in a live imaging system (Incucyte). CgRd and DRI-C23041 showed concentration-dependent inhibition with IC 50 values consistent with the previous assay ( Figure 7 ), whereas the negative control sunset yellow (SY FD C #6) showed no significant effect.

    Journal: ACS Infectious Diseases

    Article Title: Small-Molecule Inhibitors of the Coronavirus Spike: ACE2 Protein–Protein Interaction as Blockers of Viral Attachment and Entry for SARS-CoV-2

    doi: 10.1021/acsinfecdis.1c00070

    Figure Lengend Snippet: Concentration-dependent inhibition of SARS-CoV-2 pseudovirus (VSV-Δ G ) entry into hACE2/Furin expressing host cells by selected compounds. Entry of VSV-ΔG pseudoviruses bearing the SARS-CoV-2 S protein (plus GFP reporters) in ACE2/Furin overexpressing host cells (Vero-E6) was quantified via GFP fluorescence in a live imaging system (Incucyte). CgRd and DRI-C23041 showed concentration-dependent inhibition with IC 50 values consistent with the previous assay ( Figure 7 ), whereas the negative control sunset yellow (SY FD C #6) showed no significant effect.

    Article Snippet: Binding Assays SARS-CoV-2 S1 and RBD (cat. no. 40591-V08H and 40592-V08H), SARS-CoV S1+S2 (cat. no. 40634-V08B), HCoV-NL63 S1 (cat. no. 40600-V08H; all with His tag), and ACE2-Fc (cat. no. 10108-H05H) used in the binding assay were obtained from SinoBiological (Wayne, PA, USA).

    Techniques: Concentration Assay, Inhibition, Expressing, Fluorescence, Imaging, Negative Control

    Concentration-dependent inhibition of SARS-CoV-2-S-RBD binding to ACE2 by compounds of the present study. Concentration–response curves obtained for the inhibition of the PPI between SARS-CoV-2-RBD (His-tagged, 0.5 μg/mL) and hACE2 (Fc-conjugated, 1 μg/mL) in cell-free ELISA-type assay with dye (A) and nondye (B) compounds tested. The promiscuous PPI inhibitor erythrosine B (ErB) and the food colorant FD C yellow no. 6 (sunset yellow, SY) were included as a positive and negative controls, respectively. Data are mean ± SD from two experiments in duplicates and were fitted with standard sigmoid curves for IC 50 determination. Estimated IC 50 ’s are shown in the legend indicating that while suramin and chloroquine were completely inactive (IC 50 > 500 μM), several of our in-house compounds including organic dyes (CgRd, DV1, and others) as well as proprietary DRI-C compounds (e.g., DRI-C23041, DRI-C91005) showed promising activity, some even at submicromolar levels (IC 50

    Journal: ACS Infectious Diseases

    Article Title: Small-Molecule Inhibitors of the Coronavirus Spike: ACE2 Protein–Protein Interaction as Blockers of Viral Attachment and Entry for SARS-CoV-2

    doi: 10.1021/acsinfecdis.1c00070

    Figure Lengend Snippet: Concentration-dependent inhibition of SARS-CoV-2-S-RBD binding to ACE2 by compounds of the present study. Concentration–response curves obtained for the inhibition of the PPI between SARS-CoV-2-RBD (His-tagged, 0.5 μg/mL) and hACE2 (Fc-conjugated, 1 μg/mL) in cell-free ELISA-type assay with dye (A) and nondye (B) compounds tested. The promiscuous PPI inhibitor erythrosine B (ErB) and the food colorant FD C yellow no. 6 (sunset yellow, SY) were included as a positive and negative controls, respectively. Data are mean ± SD from two experiments in duplicates and were fitted with standard sigmoid curves for IC 50 determination. Estimated IC 50 ’s are shown in the legend indicating that while suramin and chloroquine were completely inactive (IC 50 > 500 μM), several of our in-house compounds including organic dyes (CgRd, DV1, and others) as well as proprietary DRI-C compounds (e.g., DRI-C23041, DRI-C91005) showed promising activity, some even at submicromolar levels (IC 50

    Article Snippet: Binding Assays SARS-CoV-2 S1 and RBD (cat. no. 40591-V08H and 40592-V08H), SARS-CoV S1+S2 (cat. no. 40634-V08B), HCoV-NL63 S1 (cat. no. 40600-V08H; all with His tag), and ACE2-Fc (cat. no. 10108-H05H) used in the binding assay were obtained from SinoBiological (Wayne, PA, USA).

    Techniques: Concentration Assay, Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay

    Identification of the binding partner by protein thermal shift. Differential scanning fluorimetry assay indicating SARS-CoV-2 RBD and not ACE2 as the binding partner of the present SMI compounds. The presence of Congo red (top) or DRI-C23041 (bottom) at 10 μM caused clear shifts in the melting temperature of the protein for RBD as indicated by the derivatives d F /d T (left; purple vs blue line), but not for hACE2 (right) (smaller insets are normalized fluorescence F data).

    Journal: ACS Infectious Diseases

    Article Title: Small-Molecule Inhibitors of the Coronavirus Spike: ACE2 Protein–Protein Interaction as Blockers of Viral Attachment and Entry for SARS-CoV-2

    doi: 10.1021/acsinfecdis.1c00070

    Figure Lengend Snippet: Identification of the binding partner by protein thermal shift. Differential scanning fluorimetry assay indicating SARS-CoV-2 RBD and not ACE2 as the binding partner of the present SMI compounds. The presence of Congo red (top) or DRI-C23041 (bottom) at 10 μM caused clear shifts in the melting temperature of the protein for RBD as indicated by the derivatives d F /d T (left; purple vs blue line), but not for hACE2 (right) (smaller insets are normalized fluorescence F data).

    Article Snippet: Binding Assays SARS-CoV-2 S1 and RBD (cat. no. 40591-V08H and 40592-V08H), SARS-CoV S1+S2 (cat. no. 40634-V08B), HCoV-NL63 S1 (cat. no. 40600-V08H; all with His tag), and ACE2-Fc (cat. no. 10108-H05H) used in the binding assay were obtained from SinoBiological (Wayne, PA, USA).

    Techniques: Binding Assay, Fluorimetry Assay, Fluorescence

    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