Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Key resources and reagents

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Blocking Assay

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Key resources and reagents for the SB system

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques:

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Key resources and reagents for the ANG system

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques:

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Analyses of CA-p24 antigen concentration on diverse HIV isolates. Twenty-two HIV-1 isolates were tested through ABR-, ANG-, SB-, and RND-CA-p24 ELISAs for detection of CA-p24 antigen. All ELISAs detected most of the isolates, except RND-CA-p24 ELISA which did not detect the 92UG029 isolate. PBS was used as negative control. Values were measured in two independent ELISA runs and error bars represent the standard deviation (SD). The CA-p24 axis is log-scaled in the graph. The list of HIV isolates can be found in Table . ABR = Aalto Bio Reagents; ANG = Anogen; SB = Sino Biological; RND = R&D Systems

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Negative Control, Standard Deviation

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Fold-change in CA-p24 detection of HIV-1 A/B isolates relative to ABR-CA-p24 ELISA. ANG-CA-p24 ELISA exhibits low reactivity for HIV-1 A and B subtypes when compared to ABR-CA-p24 ELISA. RND-CA-p24 ELISA shows acceptable reactivity for HIV-1 A and B subtypes when compared to ABR-CA-p24 ELISA, but low reactivity for the SI22 isolate. SB-CA-p24 ELISA displays good reactivity for HIV-1 A and B subtypes when compared to ABR-CA-p24 ELISA, but low reactivity for the LAI isolate. ABR = Aalto Bio Reagents; ANG = Anogen; SB = Sino Biological; RND = R&D Systems

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Virology Journal

Article Title: In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

doi: 10.1186/s12985-023-02242-5

Figure Lengend Snippet: Comparative reactivity towards HIV-1 isolates between assays as determined by Pearson’s r correlation. ANG-, SB-, and RND-CA-p24 ELISAs exhibit a significantly high correlation with ABR-CA-p24 ELISA. ANG-CA-p24 ELISA exhibits a significantly high correlation with RND-CA-p24 ELISA, whereas both RND- and ANG-CA-p24 ELISA show a low correlation with SB-CA-p24 ELISA. Concentrations of CA-p24 are displayed in ng/mL. ABR = Aalto Bio Reagents; ANG = Anogen; SB = Sino Biological; RND = R&D Systems

Article Snippet: Antibodies , Mouse Anti-HIV-CA-p24 (capture) , Sino Biological , 11695-MM08.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Virologica Sinica

Article Title: Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

doi: 10.1016/j.virs.2023.06.005

Figure Lengend Snippet: Expression and S1/S2 cleavage efficiency of SARS-CoV-2 variants' spike proteins. A Expression of SARS-CoV-2 variants' spike proteins in 293T cells. mCherry fluorescence intensity indicates spike protein expression, and GFP fluorescence intensity is the reference control. The fluorescence intensity ratio of mCherry/GFP indicates the relative expression level. The Y axis was normalized with S-WT fluorescence intensity ratio value. The numbers labeled on the bars indicate the average fold change of variants' spike protein expression relative to the S-WT. Omicron indicates BA.1 sublineage (also termed B.1.1.529) here. B , C Spike protein cleavage of pseudovirus (PVs) of the WT and variants probed with antibody against RBD. HIV-1 p24 was used to normalize the protein sample loading. D , E S1/S2 cleavage efficiency of SARS-CoV-2 spikes. The spike proteins of WT and variants were expressed in HEK293T cells and at 48 ​h post-transfection, cell lysates were made for Western blot analysis using the antibody against RBD. GAPDH served as the loading control. Full-length spike (S0) and S1 proteins are indicated. The ratio of S1 to the total spike was quantitatively analyzed using ImageJ. Data represent the mean ​± ​SD of 3–5 independent replicates. Significance was analyzed by one-way ANOVA. P values less than 0.05 were considered to be statistically significant. ∗∗∗∗, P ​< ​0.0001; ∗∗∗, P ​< ​0.001; ∗∗, P ​< ​0.01; ∗, P ​< ​0.05; ns, not significant ( P ​> ​0.05).

Article Snippet: The following primary antibodies were used: mouse anti-SARS-CoV-2 spike protein antibody (Sino Biological, Cat: 40592-MM117), mouse anti-HIV-1 p24 protein monoclonal antibody (Sino Biological, Cat: 11695-MM15), and mouse anti-GAPDH antibody (Abcam, ab8245).

Techniques: Expressing, Fluorescence, Control, Labeling, Transfection, Western Blot

Journal: Virologica Sinica

Article Title: Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

doi: 10.1016/j.virs.2023.06.005

Figure Lengend Snippet: Cell entry ability of SARS-CoV-2 pseudotyped variants bearing mutate spike proteins. A The ability of SARS-CoV-2 pseudotyped variants entering into 293T-hACE2 cells (only expressing hACE2), 293T-hACE2-TMPRSS2 cells (co-expressing hACE2 and TMPRSS2), Caco-2 ​cells (co-expressing hACE2 and TMPRSS2) and 293T cells (expressing low hACE2). B Entry comparison of pseudotyped variants in cells overexpressing hACE2 and TMPRSS2 or without TMPRSS2. HIV-1 p24 was used to normalize the PVs' additive does, and the data was calculated at 5 ​ng p24. The numbers labeled on the bars indicate ( A ) the average fold change of PV variants relative to the PV-WT or ( B ) the average fold change of 293T-hACE2-TMPRSS2 cells relative to 293T-hACE2 cells. Data are mean ​± ​SD of 3 independent replicates. Mock, uninfected control (no virus); P values less than 0.05 were considered to be statistically significant. ∗∗∗∗, P ​< ​0.0001; ∗∗∗, P ​< ​0.001; ∗∗, P ​< ​0.01; ∗, P ​< ​0.05; ns, not significant ( P ​> ​0.05). Significance was analyzed by ( A ) one-way ANOVA or ( B ) t -test.

Article Snippet: The following primary antibodies were used: mouse anti-SARS-CoV-2 spike protein antibody (Sino Biological, Cat: 40592-MM117), mouse anti-HIV-1 p24 protein monoclonal antibody (Sino Biological, Cat: 11695-MM15), and mouse anti-GAPDH antibody (Abcam, ab8245).

Techniques: Expressing, Comparison, Labeling, Control, Virus

Journal: Virologica Sinica

Article Title: Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

doi: 10.1016/j.virs.2023.06.005

Figure Lengend Snippet: Cross-species infection of SARS-CoV-2 pseudotyped variants bearing mutate spike proteins. A The ability of SARS-CoV-2 pseudotyped variants entering into cells expressing ACE2 orthologs of human, mouse, mouse, rat, rhesus, Odocoileus virginianus and with TMPRSS2 (blue columns) or without (orange columns). B A heatmap shows the fold change of the cross-species infection performance of PV variants. The Control group is the ability of PV-WT to enter into 293T-hACE2 cells. The color bar represents the scale. HIV-1 p24 was used to normalize the PVs' additive does, and the data was calculated at 5 ​ng p24. Data are mean ​± ​SD of 3–4 replicates (four for Mu, and three for others) and were normalized to the WT of the individual experiment. Mock, uninfected control (no virus); P values less than 0.05 were considered to be statistically significant. ∗∗∗∗, P ​< ​0.0001; ∗∗∗, P ​< ​0.001; ∗∗, P ​< ​0.01; ∗, P ​< ​0.05; ns, not significant ( P ​> ​0.05). Significance was analyzed by one-way ANOVA.

Article Snippet: The following primary antibodies were used: mouse anti-SARS-CoV-2 spike protein antibody (Sino Biological, Cat: 40592-MM117), mouse anti-HIV-1 p24 protein monoclonal antibody (Sino Biological, Cat: 11695-MM15), and mouse anti-GAPDH antibody (Abcam, ab8245).

Techniques: Infection, Expressing, Control, Virus

Journal: Journal of Virology

Article Title: The “LLQY” Motif on SARS-CoV-2 Spike Protein Affects S Incorporation into Virus Particles

doi: 10.1128/jvi.01897-21

Figure Lengend Snippet: Y756 mutation attenuates S protein synthesis and the packaging of S-enveloped pseudotyped virus. (A) Western blot analysis of the expression and processing of S or its mutants in the pseudoviruses producer cells at 48 h after transfection with the indicated antibodies. HIV p24 and GAPDH were used as a loading control for the cell lysate. (B and C) Western blot analysis of the S protein or its mutants in ultracentrifuge pellets from cell culture supernatant with the indicated primary antibodies on the left blots. Uncleaved S and its cleaved S1or S2 products, as well as the HIV p24 and β-actin proteins, are also indicated on the left. The asterisk indicates S protein polymer. (D) Electron microscopy of the pseudotyped virus particles. The viral particles in supernatants were purified, concentrated, and added to 400-mesh copper grids covered with carbon-coated collodion film and then stained with phosphotungstic acid (1.0% [wt/vol]) for imaging using electron microscopy. (E and F) Infection analysis of pseudoviruses in 293T-hACE2 cells was performed by continuous gradient dilution and luciferase activity analysis. Data are presented as means ± SD from 6 replicates. *** * , P < 0.0001 (Student t test). Data are representative of those from three independent experiments.

Article Snippet: Rabbit monoclonal antibody against the SARS-CoV-2 receptor-binding domain (RBD) (no. 40592-R001), rabbit polyclonal antibody against SARS-CoV-2 nucleocapsid (no. 40588-T62) and spike S2 (no. 40590-T62), and HIV-1 p24 protein (group M, subtype B, strain 92418) antibody (no. 11695-T48) were purchased from Sino Biological (China).

Techniques: Mutagenesis, Western Blot, Expressing, Transfection, Cell Culture, Electron Microscopy, Purification, Staining, Imaging, Infection, Luciferase, Activity Assay

Journal: Journal of Virology

Article Title: The “LLQY” Motif on SARS-CoV-2 Spike Protein Affects S Incorporation into Virus Particles

doi: 10.1128/jvi.01897-21

Figure Lengend Snippet: L753, F759, or T998 mutation interferes with the packaging of S-enveloped pseudotyped viruses. (A) Western blot analysis of the expression and processing of WT S or the L753G, F759G, or T998G mutant in the pseudoviruses producer cells at 48 h after transfection with the indicated antibodies. HIV p24 and β-actin were used as a loading control for the cell lysate. (B) Western blot analysis of the S protein or its mutants in ultracentrifuge pellets from cell culture supernatant by the indicated primary antibodies. (C) Relative quantification by grayscale scanning of the full-length S or cleaved S2 products from Western blot (B) with anti-S2 antibody. HIV p24 was used as a loading control. (D) Infection analysis of pseudoviruses with WT S or the L753G, F759G, or T998G mutant in 293T-hACE2 cells was performed by continuous gradient dilution and luciferase activity analysis. Data are presented as means ± SD from 6 replicates. *** * , P < 0.0001 (Student t test). Data are representative of those from three independent experiments.

Article Snippet: Rabbit monoclonal antibody against the SARS-CoV-2 receptor-binding domain (RBD) (no. 40592-R001), rabbit polyclonal antibody against SARS-CoV-2 nucleocapsid (no. 40588-T62) and spike S2 (no. 40590-T62), and HIV-1 p24 protein (group M, subtype B, strain 92418) antibody (no. 11695-T48) were purchased from Sino Biological (China).

Techniques: Mutagenesis, Western Blot, Expressing, Transfection, Cell Culture, Infection, Luciferase, Activity Assay

Journal: iScience

Article Title: Enhanced reverse zoonotic potential and immune evasion by omicron JN.1 variant

doi: 10.1016/j.isci.2025.112824

Figure Lengend Snippet: Transduction of various omicron S pseudovirions on 293/hACE2 and Calu3 cells (A) S proteins incorporation in pseudovirions. Pseudovirions with different omicron S proteins were pelleted down by centrifugation through 20% sucrose cushion and separated in a 10% SDS-PAGE. Detection of S proteins in pseudovirions was performed by Western blot using rabbit polyclonal anti-S2 antibodies. The p24 served as the loading controls. Experiments were done three times and one representative was shown. (B and C) Entry of pseudovirions of omicron variants. HEK 293/hACE2 and Calu3 cells were transduced with omicron S pseudovirions and lysed at 40 h post-transduction. The transduction efficiencies were determined according to luciferase activities. Experiments were done three times in triplicate and one representative is shown. The statistical difference relative to WT was determined by one-way ANOVA with Dunnett’s multiple testing correction ( n = 3). The statistical difference between BA.2.86 and JN.1 was determined by a two-tailed t-test ( n = 3). Error bars indicate SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Rabbit polyclonal against SARS-CoV-2 S2 antibodies (Cat#40509-T62), rabbit polyclonal against HIV-1 Gag-p24 antibodies (Cat#11695-T62), mouse polyclonal against FLAG tag antibodies (Cat#109143-MM13) and mouse polyclonal against beta-actin antibodies (Cat#100166-MM10) for western blotting and chimeric monoclonal against SARS-CoV-2 S2 antibody (Cat#40590-D001) used for flow cytometry was purchased from SinoBiological Inc (Beijing, China).

Techniques: Transduction, Centrifugation, SDS Page, Western Blot, Luciferase, Two Tailed Test