Simple immunoaffinity method to purify recombinant hepatitis B surface antigen secreted by transfected mammalian cells

J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Apr 25;787(2):303-11. doi: 10.1016/s1570-0232(02)00954-6.

Abstract

Purification of recombinant hepatitis B surface antigen (recHBsAg) produced in a stable Chinese hamster ovary (CHO) cell line was evaluated using Linx Affinity Purification System (Invitrogen, USA). To purify HBsAg secreted by this cell line, a murine monoclonal antibody (MAbAH1) raised against native HBsAg was used. The purified AH1MAb was conjugated with phenyldiboronic acid (PDBA) and immobilized on the immunoaffinity chromatographic support. Using an optimized protocol the affinity column was able to purify recHBsAg from supernatant of mammalian cells cultures with more than 80% purity. This method showed to be simple and quicker than the current ultracentrifugation methods. The method is also efficient and economical in obtaining purified recHBsAg.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Chromatography, Affinity / methods*
  • Cricetinae
  • Electrophoresis, Polyacrylamide Gel
  • Hepatitis B Surface Antigens / isolation & purification*
  • Recombinant Proteins / isolation & purification
  • Transfection

Substances

  • Hepatitis B Surface Antigens
  • Recombinant Proteins