Purification of retinal S-antigen to homogeneity by the criterion of gel electrophoresis silver staining

Abstract

This report describes a procedure by which high performance liquid chromatographic (HPLC) techniques are used to obtain homogeneous S-antigen. Conventionally prepared S-antigen was purified further by fractionation on the anion exchange Mono-Q column followed by gel filtration on a TSK-3000 column. This procedure produced an S-antigen preparation, which appeared homogeneous by gel electrophoresis using the very sensitive silver staining method. The purified material retained its immunochemical and uveitogenic activity. The availability of homogeneous S-antigen will facilitate studies aimed at elucidating, at the molecular level, the mechanism by which S-antigen induces uveitis.