大肠杆菌可溶性表达人鳞状上皮细胞癌抗原的制备及应用

doi:10.13560/j.cnki.biotech.bull.1985.2017-0228

生物技术通报 ›› 2017, Vol. 33 ›› Issue (9): 252-258.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0228

大肠杆菌可溶性表达人鳞状上皮细胞癌抗原的制备及应用

陈春野¹,刘剑¹,朱瑞¹,李姝璇¹,叶江辉¹,王玮¹,潘德全¹,徐飞海²,程通¹,夏宁邵¹

1. 厦门大学公共卫生学院分子疫苗学与分子诊断学国家重点实验室国家传染病诊断试剂与疫苗工程技术研究中心,厦门 361102；
2. 厦门万泰凯瑞生物技术有限公司,厦门 361003

收稿日期:2017-03-22 出版日期:2017-09-01 发布日期:2017-09-15
作者简介:陈春野,男,硕士研究生,研究方向：转化医学；E-mail：80911843@qq.com
基金资助:
国家高技术研究发展计划（“863”计划）（2011AA02A101）

Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

CHEN Chun-ye¹,LIU Jian¹,ZHU Rui¹,LI Shu-xuan¹,YE Jiang-hui¹,WANG Wei¹,PAN De-quan¹,XU Fei-hai²,CHENG Tong¹,XIA Ning-shao¹

1. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics,National Institute of Diagnostics and Vaccine Development of Infectious Disease,School of Life Science,Xiamen University,Xiamen 361102；
2. Xiamen WANTAI Inndox Biotechnology Co.,Ltd., Xiamen 361003

Received:2017-03-22 Published:2017-09-01 Online:2017-09-15

摘要/Abstract

摘要： 旨在建立基于大肠杆菌表达系统的高效可溶性表达人鳞状上皮细胞癌抗原（SCCAg）方法,获得具有较好活性的重组SCCAg抗原并应用于建立抗原检测方法。基于pGEX-6P-1载体和大肠杆菌E. coli ER2566菌株开展重组SCCAg抗原可溶性表达纯化方法研究,评价纯化抗原活性,筛选特异性单克隆抗体,初步建立并评价SCCAg抗原检测方法。结果显示,pGEX-6P-1载体和E. coli ER2566菌株可用于建立较高效的可溶性表达和纯化SCCAg抗原的方法,获得了具有较高纯度和活性的重组SCCAg抗原,筛选获得特异性单克隆抗体并初步建立了SCCAg管式化学发光检测方法。建立了有效的基于大肠杆菌表达系统的可溶性表达和纯化SCCAg的方法。

关键词: 人鳞状上皮细胞癌抗原, 大肠杆菌表达系统, 可溶性表达, 单克隆抗体

Abstract: The aims of this study are to establish a method for efficient soluble expression of human squamous cell carcinoma antigen（SCCAg）based on Escherichia coli expression system and obtain the recombinant SCCAg antigen in fine activity,then apply it in the detection method establishment of antigen. The study on the method of soluble expression and purification of recombinant SCCAg antigen was conducted based on pGEX-6P-1 vector and E. coli ER2566 strain. The activity of the purified antigen was evaluated by Abbott Kit and the specific monoclonal antibody was screened by indirect ELISA. It was proved that PGEX-6P-1 vector and E. coli strain ER2566 could be used to establish efficient soluble expression and purification method for recombinant SCCAg antigen. Moreover,the recombinant SCCAg antigen was proved to be in high purity and activity. Thus,the SCCAg detection method of chemical luminous tube was established with the specific monoclonal antibodies. In conclusion,an effective method for the expression and purification of SCCAg,which is based on the E. coli expression system,is established.

Key words: human squamous cell carcinoma antigen, Escherichia coli expression system, soluble expression, monoclonal antibody

陈春野,刘剑,朱瑞,李姝璇,叶江辉,王玮,潘德全,徐飞海,程通,夏宁邵. 大肠杆菌可溶性表达人鳞状上皮细胞癌抗原的制备及应用[J]. 生物技术通报, 2017, 33(9): 252-258.

CHEN Chun-ye,LIU Jian,ZHU Rui,LI Shu-xuan,YE Jiang-hui,WANG Wei,PAN De-quan,XU Fei-hai,CHENG Tong,XIA Ning-shao. Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli[J]. Biotechnology Bulletin, 2017, 33(9): 252-258.

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大肠杆菌可溶性表达人鳞状上皮细胞癌抗原的制备及应用

Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

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