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Physicochemical properties of recombinant hepatitis B surface antigen expressed in mammalian cell (C127)

  • Young Soo Lee
  • Byong Kak Kim
  • Eung-Chil Choi
Research Articles

Abstract

The physicochemical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10–15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAG by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCl density gradient method was 1.19 g/ml and the isoelectric point by Mono PTM HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. Fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and its major component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derived r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

Key words

Recombinant hepatitis B surface antigen Physicochemical properties Mouse fibroblast cell (C127) Fluorophore-assisted carbohydrate electrophoresis 

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Copyright information

© The Pharmaceutical Society of Korea 1998

Authors and Affiliations

  • Young Soo Lee
    • 1
  • Byong Kak Kim
    • 1
  • Eung-Chil Choi
    • 1
  1. 1.College of PharmacySeoul National UniversitySeoulKorea

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