[Expression and purification of IFNbeta-HSA fusion protein in Pichia pastoris].
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.
Zhang
Qi
Q
Department of Cellular and Molecular Pharmacology, School of Medicine and Pharmacology, Jiangnan University, Wuxi 214122, China.
Lei
Jianyong
J
Ding
Yuedi
Y
Chen
Yun
Y
Qu
Lin
L
Chen
Shuxian
S
Jin
Jian
J
D005285:Fermentation; D006801:Humans; D016899:Interferon-beta / Q000096:biosynthesis* / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D012709:Serum Albumin / Q000096:biosynthesis* / Q000235:genetics
D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D016899:Interferon-beta