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重组人碱性成纤维细胞生长因子的柱复性研究
Study on Refolding of Recombinant Human Basic Fibroblast Growth Factor by Gel Filtration
【摘要】 收集rhbFGF工程菌菌体,包涵体纯化后经凝胶过滤柱复性,所得复性后bFGF通过肝素亲和柱纯化并测活。各种复性方案中复性后bFGF比活最高为2.5×105U/mg,得率最高为74%。复性液中的变性剂浓度及是否含有还原剂,柱流速及柱体积都会影响复性后bFGF的得率和活性。
【Abstract】 After rhbFGF producing BL21(DE3) bacteria were collected, the inclusion bodies were extracted and refolded by gel filtration. The refolded rhbFGF was purified by affinity chromatography and the bioactivities of rhbFGF were detected by MTT assay. rhbFGF was obtained at most with specific activity of 2.5×105U/mg and the yield of 74 % in one of the refolding projects. The refolding of rhbFGF was effected by the concentration of denaturent in refolding solution, flow rate and volume of column, 0.1mol/L 2-ME in refolding solution proved to be helpful.
【基金】 国家“十五”重大专项(2002AA2Z3344)
- 【文献出处】 中国生物工程杂志 ,Progress In Biotechnology , 编辑部邮箱 ,2005年06期
- 【分类号】R346
- 【被引频次】2
- 【下载频次】156