AIB1蛋白双单克隆抗体夹心ELISA检测法的建立及初步应用

Estabilishment of double-antibody sandwich ELISA to detect AIB1 protein and preliminary application

目的建立双单克隆抗体夹心ELISA方法检测血清中AIB1的水平,探讨其在AIB1蛋白检测中的应用价值。方法应用抗AIB1-C单克隆抗体4B9F12作为固相抗体,Biotin-1A2E1作为标记抗体,ELISA双抗体夹心法检测AIB1。构建及优化的内容包括:最适包被浓度、最适包被缓冲液、生物素标记抗体工作的浓度;鉴定的指标包括:敏感性、特异性、稳定性等。结果使用0.05 mo.lL-1碳酸盐缓冲液(pH9.6)为包被缓冲液,包被抗体4B9F12工作浓度为10μg.mL-1,Biotin-1A2E1工作浓度选择1:500;采用双抗体夹心法检测外周血中AIB1的特异性和敏感性均较高。结论抗AIB1双抗体夹心ELISA方案的构建、鉴定达到了预期目的 ,为临床研究奠定了基础。

OBJECTIVE To develop the Sandwich ELISA kit for detection of AIB1,and explore its value of detection to AIB1 protein.METHODS The Sandwich ELISA kit was based on two strain McAbs,1A2E1 and 4B9F12.1A2E1 was biotinylated and acted as labeling antibody.4B9F12 was acted as immobilizing McAb.All experimental data were to set up Sandwich ELISA kit and to find out the most suitable reaction system.RESULTS The concentration of the immobilized monoclonal antibody was 10 μg·mL-1 in 0.01 mol·L-1 pH9.6 CB,and working co...