Expression and purification of IFNbeta-HSA fusion protein in Pichia pastoris.
- Author:
Qi ZHANG
1
;
Jianyong LEI
;
Yuedi DING
;
Yun CHEN
;
Lin QU
;
Shuxian CHEN
;
Jian JIN
Author Information
1. Department of Cellular and Molecular Pharmacology, School of Medicine and Pharmacology, Jiangnan University, Wuxi 214122, China.
- Publication Type:Journal Article
- MeSH:
Fermentation;
Humans;
Interferon-beta;
biosynthesis;
genetics;
Pichia;
genetics;
metabolism;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
isolation & purification;
Serum Albumin;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2009;25(11):1746-1752
- CountryChina
- Language:Chinese
-
Abstract:
In order to obtain enough fusion protein for developing preclinical studies of IFNbeta-HAS, we screened Pichia pastoris transformants expressing high-level protein by immunology method. The yield of IFNbeta-HSA was about 500 mg/L by fed-batch fermentation. The purity of IFNbeta-HSA reached 96% through the steps of ultrafiltration, Blue Sepharose FF, Ni2+-IMAC and DEAE Sepharose FF. Analysis of Western blotting showed that IFNbeta-HSA had the antigenicity of IFNbeta and HSA. The specific activity was about 1.96 x 10(7) IU/mg by standard survival activity test on WISH cells challenged with VSV virus. This study provided a method to produce IFNbeta-HSA.
