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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.coliER2566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.coliER2566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。
The aim of the present study was to establish a highly efficient and soluble method for expressing human SCCAg based on the expression system of Escherichia coli to obtain recombinant SCCAg antigen with good activity and to establish an antigen detection method. Based on the pGEX-6P-1 vector and E.coliER2566 strain, the recombinant expression vector of SCCAg antigen was purified and purified. The activity of purified antigen was evaluated and the specific monoclonal antibodies were screened. The detection method of SCCAg antigen was initially established and evaluated. The results showed that pGEX-6P-1 vector and E.coliER2566 strain could be used to establish a more efficient method of soluble expression and purification of SCCAg antigen. The recombinant SCCAg antigen with higher purity and activity was obtained and the specific monoclonal antibody was screened The SCCAg tube chemiluminescence detection method was initially established. An efficient method to express and purify SCCAg based on E.coli expression system was established.