重组人α2b型干扰素的纯化与鉴定  

Purification and Characterization of Recombinant Human Interferon-α2b

作　　者：周圆[1] 金冬雁[1] 曾庆[1] 李玉英[1] 迟捷 侯云德[1] 

机构地区：[1]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室,北京100052

出　　处：《生物工程学报》1994年第1期61-65,共5页Chinese Journal of Biotechnology

基　　金：本工作为国家863高技术研究发展计划生物领域资助项目;已于1992年5月通过专家鉴定

摘　　要：采用单克隆抗体亲和层析一步纯化法对大肠杆菌表达的重组人ａ２ｂ型干扰素进行了纯化，然后利用凝胶过滤高压液相层析、ＳＤＳ－聚丙烯酰胺凝胶电泳和Ｎ端２５个氨基酸的序列测定等方法对纯化产品进行了鉴定。表明一步纯化的ＩＦＮ－ａ２ｂ达到９５％以上的统一计划，其比活性为２．５４×１０×８ｕ／ｍｇ．但重组ＩＦＮ－ａ２ｂ产品Ｎ端不均一，含有约３０％的去Ｍｅｔ分子和约７０％的带Ｍｅｔ分子。从蛋白质序列的水平上证实。Recombinant human α2b-type interferon (IFN-α2b) expressed in E. coli was purified to >95% homogeneity through affinity chromatography on Sepharose 4B coupled with anti-IFN-α2a monoclonal antibody. The purified protein, with a specific an-tiviral activity of 2. 54×108 IU/mg protein, appeared as a single band on the SDS-PAGE electrophoretogram and a single peak on the HPLC chromatogram. The N- terminal 25 amino-acid sequence of recombinant IFN-α2b was determined on Applied Biosystems 477A protein/peptide sequencer and was shown to be correct. The purified protein was found to be heterogenous, comprising two sequences which differ merely by the initiating Met 70% of Met-plus speicies and 30% of Met-minus species.

关 键 词：a2b型 干扰素 提纯 鉴定 重组 

分 类 号：R394.8[医药卫生—医学遗传学]