人β干扰素与人血清白蛋白融合蛋白的表达及纯化  

Expression and purification of IFNβ-HSA fusion protein in Pichia pastoris

作　　者：张琦 雷楗勇 丁月娣 陈蕴 屈琳 陈淑娴 金坚 

机构地区：[1]江南大学医药学院细胞与分子药理学实验室,无锡214122

出　　处：《生物工程学报》2009年第11期1746-1752,共7页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划(863计划)(No.2006AA02Z153)资助~~

摘　　要：为开展人β干扰素-人血清白蛋白融合蛋白(IFNβ-HAS)的临床前研究,本研究采用免疫学方法对毕赤酵母转化子进行筛选,得到高产菌株8株。在5L发酵罐上对高产菌株进行诱导表达,产量达500mg/L。发酵液经超滤浓缩、染料亲和层析、镍离子螯合亲和层析和DEAE离子交换层析纯化后,融合蛋白纯度达到96%。Westernblotting杂交表明融合蛋白具有人血清白蛋白和人β干扰素的抗原性,细胞病变抑制法测定IFNβ-HSA比活性为1.96×107IU/mg。建立了融合蛋白的生产工艺,为IFNβ-HSA的临床前研究奠定了基础。In order to obtain enough fusion protein for developing preclinical studies of IFNβ-HAS,we screened Pichia pastoris transformants expressing high-level protein by immunology method.The yield of IFNβ-HSA was about 500 mg/L by fed-batch fermentation.The purity of IFNβ-HSA reached 96% through the steps of ultrafiltration,Blue Sepharose FF,Ni2＋-IMAC and DEAE Sepharose FF.Analysis of Western blotting showed that IFNβ-HSA had the antigenicity of IFNβ and HSA.The specific activity was about 1.96×107 IU/mg by standard survival activity test on WISH cells challenged with VSV virus.This study provided a method to produce IFNβ-HSA.

关 键 词：Β干扰素 融合蛋白 毕赤酵母 

分 类 号：Q78[生物学—分子生物学]