Fig. 7.

Absolute quantification of serum antigen-specific antibody titers

(A-B) Absolute quantification of NP-specific antibody concentration in serum. C57BL/6 mice were immunized with the alum-precipitated NP-CGG. ELISA plates were coated with NP₂₁-BSA (A) or NP₂-BSA (B). Antibodies bound to the plates were detected using HRP-conjugated anti-mouse isotype-specific antibodies. NP3S2-IgG1 mAb from Fig. 3 or NP3S2-IgM, IgG3, IgG1, IgG2b, IgG2c, IgE, and IgA mAbs developed from NP3S2-IgG1, which are consisted of identical V-domain recognizing NP, were used as the standard. From the four-parameter logistic (4 PL) curve fit of NP3S2-derived mAbs on Prism, NP-specific antibody concentration of each sample was determined. n.d., not detectable.

(C–D) Post-immunization NP₂/NP₂₁ ratios in IgG1 (C), IgM (D), and IgG2b (D). The NP₂/NP₂₁ ratios were calculated based on the NP₂-binding antibody concentration and the NP₂₁-binding antibody concentration. Relative NP₂/NP₂₁ ratios were calculated by dividing the ratio at each time point by the average ratio at day 10. avg., average.