The purification of HBV full-length PreS protein in Pichia pastoris.
- Author:
Xue HAN
1
;
Lin-Bai YE
;
Bao-Zong LI
;
Ying-Long SHE
;
Li YE
;
Hong ZHENG
;
Bo GAO
;
Jin-Rong GAO
;
Zheng-Hui WU
Author Information
1. National Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
- Publication Type:Journal Article
- MeSH:
Hepatitis B Surface Antigens;
biosynthesis;
genetics;
Hepatitis B virus;
immunology;
Humans;
Pichia;
genetics;
metabolism;
Protein Precursors;
biosynthesis;
genetics;
Recombinant Proteins;
biosynthesis;
genetics;
immunology;
isolation & purification;
Viral Envelope Proteins;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(5):708-712
- CountryChina
- Language:Chinese
-
Abstract:
The Pichia pastoris strain GS115-PreS could produce a high expression level of full-length PreS protein that secreted to the supernatant after methanol induction in the fermentation. The Western blot analysis showed a single band with expected molecular mass of 48kD and that the major component of the particles was the full-length PreS protein (PreS1 + PreS2 + S) and small envelope protein (S) of 48 and 28 kD, respectively. Electron microscopy image showed PreS particles with 30 nm in diameter. The supernatants of the fermentation were desalted and concentrated. Purified PreS protein was obtained by DEAE-SFF anion exchange column chromatography and the PreS particles were obtained by ultracentrifugation and sucrose density gradient. The ELISA assay results proved that both full-length PreS protein and particles showed high immunogenicity and specificity. P/N ratio further demonstrated that the immunogenicity of the particles is higher than the full-length PreS protein.
