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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

作者： Zhang, Gaiping 1 ; Xi, Jun 1 ; Wang, Xuannian 3 ; Guo, Junqing 4 ; Zhang, Hua 1 ; Yang, Yanyan 1 ; Qiao, Songlin 1 ; Wang 1 ;

作者机构： 1.Henan Acad Agr Sci, Henan Prov Key Lab Anim Immunol, Zhengzhou 450002, Peoples R China

2.Jilin Univ, Coll Anim Sci & Vet Med, Changchun 130062, Peoples R China

3.Henan Inst Sci & Technol, Coll Anim & Vet Med, Xinxiang 453003, Peoples R China

4.Zhejiang Univ, Coll Anim Sci,

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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[1]Expression, purification and characterization of a functional extracellular domain of porcine Fc gamma RII. Tian, Xiaohui,Qiao, Songlin,Zhang, Gaiping,Xi, Jun,Liu, Yunchao,Tian, Xiaohui,Wang, Aiping,Li, Xuewu,Xu, Yuqing. 2009

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