Journal: Vaccines

Article Title: Mimicking Native Display of CD0873 on Liposomes Augments Its Potency as an Oral Vaccine against Clostridioides difficile

doi: 10.3390/vaccines9121453

Figure Lengend Snippet: Immune responses in hamsters immunised orally with CD0873 alone and CD0873-MalLipo. Intestinal fluid and serum were harvested from vaccinated hamsters and from negative control hamsters and tested for the presence of mucosal IgA and serum IgG, respectively. Intestinal fluid was extracted by flushing the small intestine with 5 mL of PBS containing protease inhibitors, and serum was taken by cardiac puncture. ( A ) Intestinal fluid (5 µL) from each hamster was first fractionated by SDS-PAGE and transferred to a PVDF membrane then probed with rabbit anti-hamster IgA antibody (1:1000). Bands were detected with anti-rabbit IgG HRP (1:1000) and TMB substrate, and band intensity was quantified using ImageJ. ( B ) Serum diluted 1:10 of CD0873- and CD0873-MalLipo-vaccinated group and naïve group tested for IgG by indirect ELISA. Goat anti-hamster IgG highly cross-adsorbed Biotin antibody (1:20,000) and Streptavidin-HRP (1:200) were used for detection. Data were obtained from two independent experiments, each with three technical replicates. Error bars represent standard error of the mean (SEM). All data were analysed by Kruskal–Wallis test followed by Dunn’s multiple comparison. Error bars represent the standard error of the mean. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Wells were washed, then incubated for 2 h at room temperature in 100 µL goat anti-hamster IgG highly cross-adsorbed Biotin antibody (Sigma Merck Group, Feltam, UK) diluted 1:20,000 in PBST, washed again, then incubated for 2 h in Streptavidin-HRP (R&D Systems, Minneapolis, MN, USA) diluted 1:200 in PBST.

Techniques: Negative Control, SDS Page, Indirect ELISA