Size exclusion chromatography of hepatitis B surface antigen particles

Summary To find the factors responsible for the broadening of the recombinant-hepatitis B surface-antigen peak in size-exclusion chromatography, the purified material was fractionated on preparative scale followed by multiple analysis of the separated fractions. The results from chromatographic analysis suggested the presence of large particle aggregates, probably tubular structures which, however, were not detected by electron microscopy. The antigen particles ranged from 16 to 32 nm in all the fractions, except two last fractions consisting of 16–24 nm particles. The relation ELISA/Lowry increased with increasing the fraction number, being a maximum in the fraction corresponding to the maximum of the chromatographic peak. Probably, the particles which are variable in size differ from each other with respect to the efficiency of protein assembly. Fractions collected in different regions of the peak were adsorbed on alum and injected in mice. The high antibody levels were produced without significant differences in immunogenicity between samples. Presented at the 21^st ISC held in Stuttgart, Germany, 15^th–20^th September, 1996.