ADSORPTION-DESORPTION OF RECOMBINANT HEPATITIS-B SURFACE-ANTIGEN (R-HBSAG) FROM P-PASTORIS ON A DIATOMACEOUS-EARTH MATRIX - OPTIMIZATION OFPARAMETERS FOR PURIFICATION

Recombinant hepatitis B surface antigen (r-HBsAg) produced in yeast is adsorbed on a diatomaceous earth matrix for purification purposes. A pH dependence in the adsorption-elution behavior was found. The capaci ty of celite (Hyflo Super Cel) for adsorbing r-HBsAg increased with de creasing pH. Nonspecific proteins were also adsorbed, but a low pH dep endence was found. Elution from the matrix was performed using a basic pH buffer, in which r-HBsAg is more specifically adsorbed/desorbed th an contaminant proteins, permitting the purification of the r-HBsAg. A pH of 4.0 was used for adsorption and pH 8.2 was used for desorption. The described protocol allows a purification factor between three- an d fivefold with respect to contaminant proteins and sixfold with respe ct to contaminant DNA. Finally, the adsorption step was successfully s caled-up for production purposes. (C) 1993 John Wiley & Sons, Inc.