Journal: Frontiers in Immunology

Article Title: Immunization with detoxified TNFα elicits neutralizing antibodies and ameliorates inflammatory shock and autoimmune arthritis in mice

doi: 10.3389/fimmu.2026.1801223

Figure Lengend Snippet: CP-m87S-NH vaccination induces antigen-specific antibodies that block mTNFα binding to TNF receptor. (A) Immunization scheme. ICR mice were immunized intramuscularly with mTNFα vaccines (2 µg antigen) on days 0 and 14, and serum was collected on day 28. (B) Anti-mTNFα IgG titer measured by ELISA. Groups included untreated (n=3), Alum/m87S-NH (n=3), 2HPQ/m87S-NH (n=3), CPQ/m87S-NH (n=3), CP/m87S-NH (n=3), and CP/mTNF-NH (n=3). (C) mTNFα-hTNFR1 binding inhibition (sera diluted 1:15 in PBS) and (D) corresponding inhibition titers. hTNFR1-coated plates were incubated with immunized serum pre-mixed with biotinylated mTNFα at 37 °C for 1 h. After transfer to the hTNFR1-coated plate and incubated for 30 minutes at 37 °C, unbound proteins were washed off with PBST. Captured biotinylated mTNFα was detected with streptavidin-HRP and TMB substrate. Lines and bar graphs show mean ± SD for n=3 mice per group. Statistical significance was determined by one-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.001.

Article Snippet: After washing with PBST, TMB substrate (Surmodics, Cat# TMBW-1000-01) was added.

Techniques: Blocking Assay, Binding Assay, Vaccines, Enzyme-linked Immunosorbent Assay, Inhibition, Incubation