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

DOI：10.3969/j.issn.1673-1689.2012.03.011

中文关键词: 人白介素2 人血清白蛋白 融合蛋白 纯化

英文关键词: human interleukin-2 human serum albumin fusion protein purification

基金项目: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)

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中文摘要:

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。

英文摘要:

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.

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