LARGE-SCALE PURIFICATION OF HEPATITIS-B SURFACE-ANTIGEN (HBSAG)

Abstract

In Oct. 1975, a specific immunization by a formalin inactivated hepatitis B vaccine was introduced to protect patients and staff members of 3 hemodialysis units of the Loire Valley (Tours, Blois, Orleans, France). After 2 yr follow-up the innocuity and efficacy of this preparation were very satisfactory in the conditions under which it was used. A method of vaccine preparation was instituted in the development of industrial batches of vaccine to be used for broad clinical trials in France. The HBsAg purification was carried out by 4 different steps including, successively, selective adsorption-desorption on colloidal silicate (Aerosil), precipitations by polyethylene glycol, gel filtration and finally zonal ultracentrifugation. Step-by-step results of the purification are presented. Up to 65% of the starting antigen was recovered at the end of the purification process. One dose of vaccine (1 ml) has a titer in HBsAg of 1/4 in counter-current electrophoresis and a protein amount of 2-10 .mu.g/ml. It contains traces of homologous serum proteins only detectable after high concentration. Purity, antigenic quality and safety of the vaccine are analyzed in regard to its use for immunization against hepatitis B in man.