title: [Expression and purification of IFNbeta-HSA fusion protein in Pichia pastoris].
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.
authors.affiliation: Department of Cellular and Molecular Pharmacology, School of Medicine and Pharmacology, Jiangnan University, Wuxi 214122, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.forename: Jianyong
authors.initials: J
authors.lastname: Lei
authors.forename: Yuedi
authors.initials: Y
authors.lastname: Ding
authors.forename: Yun
authors.initials: Y
authors.lastname: Chen
authors.forename: Lin
authors.initials: L
authors.lastname: Qu
authors.forename: Shuxian
authors.initials: S
authors.lastname: Chen
authors.forename: Jian
authors.initials: J
authors.lastname: Jin
mesh_terms: 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
chemical_list: D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D016899:Interferon-beta