Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 6. In vitro antigenicity analysis of t-M2e-t NCs using ELISA. (A) Absorbance values from an indirect ELISA performed by coating 96-well plates with a fixed amount of either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a negative control peptide (NCP). Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of M2e detection IgG antibody, which was applied at different dilutions. (B) Absorbance values from a sandwich ELISA performed by coating 96-well plates with a fixed amount of purified mouse anti-M2e IgM monoclonal antibody as capture antibody. Dilutions of analyte (either un-cross-linked t-M2e-t peptide, cross-linked t-M2e-t NCs, or a NCP) were then added to the wells. Mouse serum collected from mice vaccinated with a gold nanoparticle and M2e formulation (from another study8−10) was used as a source of primary M2e detection IgG antibody. Cartoons of each ELISA procedure are shown below their respective graphs.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Indirect ELISA, Negative Control, Formulation, Sandwich ELISA

Journal: ACS infectious diseases

Article Title: Tyrosine-Based Cross-Linking of Peptide Antigens to Generate Nanoclusters with Enhanced Immunogenicity: Demonstration Using the Conserved M2e Peptide of Influenza A.

doi: 10.1021/acsinfecdis.1c00219

Figure Lengend Snippet: Figure 7. Immune response and survival data of mice after immunization with t-M2e-t NCs. Mice were immunized thrice, once each on day 0, 21, and 42. Blood was collected and analyzed for anti-M2e antibodies. Within 1 week of the final serum collection, mice were challenged with 3× LD50 A/California/07/2009 (H1N1) and monitored for 14 days. Two antigens were used (i) 20 μg or 5 μg t-M2e-t UCP (un-cross-linked peptide) with/without 20 μg CpG, and (ii) 20 μg or 5 μg S t-M2e-t NCs (small NCs) with/without 20 μg CpG. (A−C) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t UCP as antigen. (D−F) Anti-M2e IgG, IgG1 and IgG2a titers with t-M2e-t NC as antigen. (G, H) Body weight and survival after virus challenge of t-M2e-t UCP groups. (I, J) Body weight and survival after virus challenge of t-M2e-t NP groups. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All peptides were end-terminal acetylated and amidated in order to increase peptide stability against degradation.65−67 Goat anti-mouse IgG (1030−05), goat anti-mouse IgG1 (1070−05), and goat anti-mouse IgG2a (1080−05) secondary antibodies with HRP were obtained from Southern Biotech (AL, USA).

Techniques: Virus

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BM5 by intramuscular (IM) N = 7 and Gene Gun (GG), N = 8 immunization. Antigen-specific ELISAs were performed with sera collected 8 wk after the first immunization. A : IgG1 and IgG2a results shown are mean OD + SE for IM and GG mice; B : IgG1 to IgG2a ratios are shown for mice immunized with BM5 by IM and GG routes. Values shown are mean PD +SE. The ratios in the two groups were significantly different (p < 0.05).

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BMIF by IM (N = 12) and GG (N = 9). Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG1 and IgG2a antibodies shown are mean OD + SE; B: IgG1 to IgG2a ratios are shown for mice immunized with BMIF by IM and GG routes. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BMHSP by IM (N = 12) and GG (N = 9) route. Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG1 and IgG2a results shown are mean OD +SE; B: IgG1 to IgG2a ratios are shown for mice immunized with BMHSP by IM and GG route. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: This figure shows serum IgG isotype antibody responses in mice vaccinated with BM14 by IM (N = 12) and GG (N = 9). Antigen-specific ELISAs were performed with sera collected 8 wk after immunization. A: IgG, IgG1 and IgG2a antibody results shown are mean OD + SE; B: IgG1 to IgG2a antibody ratios are shown for mice immunized with BMHSP by IM and GG routes. Values shown are means + SE. The ratios were not significantly different.

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques:

Journal: Filaria Journal

Article Title: Antibody responses to Brugia malayi antigens induced by DNA vaccination

doi: 10.1186/1475-2883-3-1

Figure Lengend Snippet: Time course of total IgG antibodies to individual recombinant B. malayi antigens after polyvalent and monovalent DNA vaccination by IM injection. ELISAs were performed with sera collected at different times after immunization. Results shown are mean OD + SE. A: IgG antibody responses to individual antigens after polyvalent DNA vaccination (5 mice). B: IgG antibody responses to individual antigens after monovalent DNA vaccination (7 mice for BM5, 12 mice for each of the other antigens).

Article Snippet: Bound antibody was detected after incubation with peroxidase-conjugated goat anti-mouse IgG, IgG1, or IgG2a antibodies (Southern Biotechnology Associates, Birmingham, AL) for 1 hr at 37°C.

Techniques: Recombinant, Injection

Journal:

Article Title: Myeloid Cell Function in MRP-14 (S100A9) Null Mice

doi: 10.1128/MCB.23.7.2564-2576.2003

Figure Lengend Snippet: MRP-14 protein is absent in MRP-14−/− monocytes and neutrophils. (A) Myeloid cells were identified by flow cytometry by staining MRP-14+/+ and MRP-14−/− bone marrow cells with MAb 7/4. (B) The MAb 7/4-positive myeloid cells (R1) from MRP-14+/+ and MRP-14−/− mice were intracellularly stained with either MAb 2B10, specific for MRP-14 (shaded histogram), or an IgG2a isotype control MAb (open histogram). (C) In MRP-14+/+ mice, two different MRP-14-positive populations (R2 and R3) can be seen that are absent in MRP-14−/− myeloid cells. These populations correspond to monocytes (R2) and neutrophils (R3), based on their differential staining with MAb Gr-1. Open histogram represents the Gr-1-negative, 7/4-negative cells. Inset numbers represent the geometric mean fluorescence of three Gr-1-positive populations.

Article Snippet: MRP-14 was detected with MAb 2B10 (2 μg/ml) and compared to control IgG2a, followed by goat anti-rat immunoglobulin-horseradish peroxidase conjugate (1:500, Southern Biotechnology).

Techniques: Flow Cytometry, Staining, Fluorescence