Figure 5.

FACS-based detection of antigen-specific antibodies using antigen displaying PHA granules. In (A), MOG-phaP granules were incubated with serial dilutions of pooled antisera from five MOG or OVA immunized mice. Granules were extensively washed and then incubated with biotinylated anti-mouse IgG, followed by PE-conjugated streptavidin. The data was depicted in a "normalized" fashion (% of Max). One representative experiment, of at least two experiments performed, is depicted. In (B), the mean channel fluorescent for each dilution of antisera from MOG or OVA immunized mice binding to MOG-phaP was measured and depicted. In (C), ELISA was performed on anti-sera from MOG- and OVA-immunized mice. Serial dilutions of the anti-sera were added to MOG- or OVA-coated wells and incubated for 30 minutes. Biotinylated anti-mouse IgG was then added to the washed wells, followed by HRP-conjugated streptavidin and TMB substrate. The optical density was read at 450 nm and the results from one representative experiment, of at least two experiments performed, are shown. In (C), data are displayed as mean ± SEM from triplicate samples.