Antibody concentration and affinity following
immunization. (a)
Timeline of the experimental setup shows subcutaneous (SC) injection
of a model vaccine containing OVA and Poly(I:C) in a gel or bolus
formulation at day 0, antibody analysis over time following a single
administration, boost with a bolus vaccine formulation at day 90,
and analysis of the immune response 15 days after the boost. (b) Serum
anti-OVA IgG1 concentrations from day 0 to day 90 after a single injection
of vaccines (n = 5 to 19; one to four independent
experiments; mean ± s.e.m.). ***p < 0.001
and ****p < 0.0001 compared to bolus, #p < 0.05, ##p <
0.005 compared to 1:5, determined by mixed-effects analysis with Tukey’s
post hoc test. Serum anti-OVA (c) IgG end point titer, (d) IgG1 concentration,
(e) IgG2b concentration, and (f) IgG2c concentration 15 days after
bolus boost on day 90 for animals receiving either bolus, 1:5 gel,
or 2:10 gel vaccines (n = 4 to 5; mean ± s.d.).
Reported p values determined by one-way ANOVA with
Tukey’s post hoc test. (g) Model comparing competitive binding
data with KD ranging from 1 to 104 nM. (h) Representative competitive binding curves for bolus,
1:5 gel, and 2:10 gel vaccine groups after the day 90 boost compared
to an mAb reference competing with the same mAb. (i) Calculated KD values from fitted binding curves for bolus,
1:5 gel, and 2:10 gel vaccine groups (n = 4; mean
± s.d.). p values determined by one-way ANOVA
with Tukey’s post hoc test.