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目的建立雌酮硫酸钠(ESS)单克隆抗体的制备方法。方法以ESS为抗原免疫Balb/c小鼠,筛选小鼠免疫方法,并通过方阵滴定法确定间接酶联免疫吸附测定(ELISA)方法;再以杂交瘤抗体技术获得抗ESS单克隆抗体细胞株。结果 ESS抗原5次以初次免疫200μg,加强免疫100μg的免疫剂量多点注射小鼠背部皮下免疫为小鼠最佳免疫方法;确定10μg·L-1为抗原包被浓度,1∶2 000的一抗血清稀释倍数;细胞融合率达90.2%,ELISA法筛选阳性率为4.4%,并筛选得到两株特异性较好的细胞株2C8和8A7。结论该研究筛选建立了ESS小鼠免疫方法及灵敏度高、特异性好的间接ELISA筛选体系,可用于ESS单克隆抗体的制备及检测,且建立了ESS单克隆抗体制备方法。
Objective To establish a method for preparing monoclonal antibody against estrone sodium sulfate (ESS). Methods Balb / c mice were immunized with ESS as antigen and the mouse immunization method was screened. The indirect enzyme-linked immunosorbent assay (ELISA) was determined by square matrix titration. The monoclonal antibody against ESS . RESULTS: The ESS antigen was immunized with 200μg primary antibody for 5 times, and the subcutaneous immunization with multiple immunized 100μg immunized mice was the best immunization method. The optimal concentration of antigen was 10μg · L-1, Antiserum dilution ratio, the cell fusion rate was 90.2%, the positive rate of screening by ELISA was 4.4%, and two specific cell lines 2C8 and 8A7 were screened. Conclusion This study established an immune method for ESS mice and an indirect ELISA screening system with high sensitivity and specificity, which can be used for the preparation and detection of ESS monoclonal antibodies. The preparation method of ESS monoclonal antibody was also established.