Expression and purification of IFNbeta-HSA fusion protein in Pichia pastoris / 生物工程学报
Qi ZHANG; Jianyong LEI; Yuedi DING; Yun CHEN; Lin QU; Shuxian CHEN; Jian JIN.
Chinese Journal of Biotechnology
; (12): 1746-1752, 2009.
Artículo
en Zh
| WPRIM | ID: wpr-296863
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.
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