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

摘要

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.%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.