Preparation of the porcine secretory component and a monoclonal antibody against this protein

文献类型： 外文期刊

第一作者： Song, Wei-Xiang

作者： Song, Wei-Xiang;Jiang, Ping;Song, Wei-Xiang;Feng, Zhi-Xin;Bai, Yun;Wang, Hai-Yan;Ishag, Hassan Zackaria Ali;Liu, Mao-Jun;Xiong, Qi-Yan;Shao, Guo-Qing;Feng, Zhi-Xin;Shao, Guo-Qing

作者机构：

关键词： Secretory component;Prokaryotic expression;Monoclonal antibody

期刊名称：PROTEIN EXPRESSION AND PURIFICATION （ 影响因子：1.65； 五年影响因子：1.548 ）

ISSN：

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收录情况： SCI

摘要： Secretory component (SC) is a component of secretory IgA that is designated sIgA to distinguish it from IgA. The monoclonal antibody (MAb) against SC has been shown to be an excellent tool for the detection of the level of sIgA and for the evaluation of the efficacy of mucosa] immunity. To prepare a monoclonal antibody against porcine SC, a recombinant porcine SC was expressed and purified. To develop this recombinant SC, the gene encoding the porcine SC was ligated into the pCold I vector. The recombinant vector was then transformed into Escherichia coil BL 21 (DE3), and gene expression was successfully induced by isopropyl-beta-D-thiogalactoside (IPTG). After affinity purification with Ni-NTA resin and gel recovery, the recombinant SC protein was used to immunize BALB/c mice. Finally, three hybridoma cell lines showing specific recognitions of both recombinant SC and native SC were used as stable secretors of MAbs against porcine SC and were confirmed to have no reaction to porcine IgA or IgG. The successful preparations of recombinant SC protein and MAbs provide valuable materials that can be used in the mucosal infection diagnosis for porcine disease and mucosal immune evaluation for porcine vaccine, respectively. (C) 2015 Elsevier Inc. All rights reserved.

分类号： Q51

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