Preparation of IgE high affinity receptor protein and its biological function identification
10.3969/j.issn.1671-8348.2018.09.004
- VernacularTitle:IgE高亲和力受体蛋白的制备及其生物功能的鉴定
- Author:
Ting XU
1
;
Qiang ZHANG
;
Helian YU
;
Shichun WANG
;
Xiaomin XU
;
Zhongmei YI
;
Tianlun JIANG
Author Information
1. 陆军军医大学西南医院输血科
- Keywords:
IgE high affinity receptor;
IgE antibody;
anti-FcεRIα autoantibody;
enzyme-linked immunosorbent assay
- From:
Chongqing Medicine
2018;47(9):1161-1164
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare human alpha segment of high affinity IgE receptor(FcεRIα)protein by genetic engineering technology and to identify its biological function for laying the foundation for further researching the role of FcεRIα in allergic disea-ses.Methods The human FcεRIα gene was obtained by the PCR based accurate synthesis(PAS)method and the prokaryotic ex-pression vector pET-28a(+)was constructed.The FcεRIα was expressed at low temperature induction and the recombinant protein was purified by His tag.The biological function of recombinant human FcεRIα protein was identified by ELISA.Results The hu-man FcεRIα gene was amplified by PAS with a size of approximately 560 bp.The pET-FcεRIα plasmid was correct through the double enzyme digestion and sequencing identification.The human FcεRIα with a molecular weight of approximately 22 000 was in-duced and purified.The recombinant human FcεRIα could effectively detect human serum anti-FcεRIα autoantibody and could com-bined with serum IgE antibodies with high efficiency.Conclusion Human FcεRIα protein is successfully prepared,which prelimina-rily has the ability for detecting the human serum anti-FcεRIα autoantibodies and IgE antibodies,and provides a favorable practical base for further study.
