High-performance size-exclusion chromatography and molar mass measurement by low-angle laser light scattering of recombinant yeast-derived human hepatitis B virus surface antigen vaccine particles.

Two kinds of recombinant yeast-derived human hepatitis B virus surface antigens (HBsAg) for use as vaccines were analysed by high-performance size-exclusion chromatography using a TSK-GEL G60000PWXL column. One consisted of the surface antigen, a glycosylated polypeptide with of about 230 amino acid residues identical with that of human hepatitis B virus in addition to lipids derived from the yeast cell. The other differed in that it consisted of a polypeptide with an additional peptide of nine amino acid residues corresponding to the pre-S2 region. Elution from the column was monitored with a low-angle laser light-scattering detector and also UV and refractive index detectors. The results obtained indicate that both of the antigen particles gave a single symmetrical peak in the elution curves, the molar masses of the particles were 5040 and 4640 kg/mol for the former and the latter, respectively, and the particle size was homogeneous for both of the HBsAg particles. The present approach seems suitable for the characterization of such antigen particles, which will be widely used as vaccines.