Adsorption–desorption of recombinant hepatitis B surface antigen (r‐HBsAg) from P. pastoris on a diatomaceous earth matrix: Optimization of parameters for purification

Abstract

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 capacity of celite (Hyflo Super Cei) for adsorbing r‐HBsAg increased with decreasing pH. Nonspecific proteins were also adsorbed, but a low pH dependence was found. Elution from the matrix was performed using a basic pH buffer, in which r‐HBsAg is more specifically adsorbed/desorbed than 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‐ and fivefold with respect to contaminant proteins and sixfold with respect to contaminant DNA. Finally, the adsorption step was successfully scaled‐up for production purposes. © 1993 John Wiley & Sons, Inc.