人白介素2-人血清白蛋白融合蛋白的纯化及活性鉴定  

Purification and Bioactivity of the Fusion Protein Human Interleukin-2/Human Serum Albumin

作　　者：李波[1] 段作营[1] 张红梅[1] 雷楗勇 陈蕴 唐瑜菁[3] 金坚 李华钟[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学医药学院,江苏无锡214122 [3]泰山医学院生命科学系,山东泰安271016

出　　处：《食品与生物技术学报》2012年第3期289-293,共5页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(30970029);江南大学教育部工业生物技术重点实验室开放课题(KLIB-KF200704)

摘　　要：采用课题组已构建的携带人白介素2-人血清白蛋白融合蛋白(rhIL-2-HSA)基因的重组毕赤酵母Pichia pastoris GS115进行rhIL-2-HSA的分离纯化研究。发酵上清液经超滤浓缩、Blue Sepharose亲和层析、Octyl Sepharose疏水层析和DEAE Sepharose离子交换层析纯化获得较高纯度的目的产物,SDS-PAGE检测为单一条带。Western Blot分析结果显示,目的产物同时具有HSA和IL-2的抗原性,为HSA和IL-2的融合分子。经CTLL-2/MTT细胞增殖法测定,纯化的融合蛋白的生物学活性为1.147×107 IU/mg。The purification of fusion protein of human interleukin-2 and human serum albumin(rhIL-2-HSA) that was expressed and secreted in fermentation broth with recombinent Pichia pastoris GS115/pPIH was reported in this paper.Highly purified rhIL-2-HSA was separated from fermentation supernatant by ultra filter,Blue Sepharose affinity chromatography,Octyl Sepharose hydrophobic chromatography,DEAE Sepharose ion exchange chromatography.The purified fusion protein showed one single band on SDS-PAGE.The purified fusion protein could react with antibodies to IL-2 and HSA in Western Blot analysis.CTLL-2/MTT cell proliferating assay showed that the special activity of rhIL-2-HSA was 1.147×107 IU/mg.

关 键 词：人白介素2 人血清白蛋白 融合蛋白 纯化 

分 类 号：R96[医药卫生—药理学]