Efficient recovery of a functional extracellular domain of bovine IgG2 Fc receptor (boFc gamma 2R) from inclusion bodies by a rapid dilution refolding system

文献类型： 外文期刊

第一作者： Zhang, Gaiping

作者： Zhang, Gaiping;Xi, Jun;Zhang, Hua;Yang, Yanyan;Qiao, Songlin;Wang, Li;Zhang, Hong;Zhang, Gaiping;Xi, Jun;He, Liyang;Wang, Xuannian;Zhu, Yancai;Guo, Junqing

作者机构：

关键词： boFc gamma 2R;extracellular domain;rapid dilution refolding

期刊名称：JOURNAL OF IMMUNOLOGICAL METHODS （ 影响因子：2.303； 五年影响因子：2.556 ）

ISSN： 0022-1759

年卷期： 2008 年 334 卷 1-2 期

页码：

收录情况： SCI

摘要： The extracellular domain of the boFc gamma 2R gene was constructed and cloned into the Escherichia coli expression vector pET-28a. The recombinant protein was expressed at high level in E. coli BL21 (DE3) and existed mainly as inclusion bodies. The inclusion bodies were solubilized in 6 M guanidine hydrochloride and refolded by rapid dilution. After renaturation, the purity of the recovered recombinant protein was up to 95%. ELISA assay showed that the renatured recombinant protein could inhibit bovine IgG2 binding to expressed boFc gamma 2R on the COS-7 cell surface with an IC50 value of 0.68 mu M. The overall yield of the active rsbo2R was up to 20 mg/l of culture. Crystals of the rsbo2R were grown at 293 K by the hanging-drop vapour diffusion method showed weak diffraction. (C) 2008 Elsevier B.V. All rights reserved.

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