Association of Normal Serum Protein Antigens with Chimpanzee Hepatitis B Surface Antigen Particles

HBsAg/adw was purified from 2.6 liters of pooled plasma from a single chimpanzee carrier by polyethylene glycol (PEG) precipitation followed by isopycnic and rate zonal centrifugation. The different morphological populatilons of HBsAg separated in the final rate zonal centrifugation step were combined into seven pools: two fractions rich in filaments and Dane particles, two pools composed of filaments and 20--28-nm spheres, and three fractions containing mostly 20--28-nm spheres. The purified preparations of HBsAg analyzed for normal serum protein contaminants revealed albumin and traces of IgG. The same samples analyzed after Tween-80 treatment, revealed enhanced quantitites of the previous two contaminants, and in addition, transferrin, traces of alpha2-macroglobulin, IgM, and complement (C3/C3c). The residual contaminants were mostly removed by further purification and fractionation after detergent treatment using zone convection electrofocusing and rate zonal centrifugation. Our findings indicate that conventional purification techniques will not provide preparations of HBsAg free of traces of serum protein contaminants. Many of these are released only by detergent treatments and subsequent purification. It is not yet clear whether detergents release these contaminants from the interior of the particles or from firm association or incorporation within the membranes.