孕激素受体A型原核表达及单克隆抗体制备

Expression of progesterone receptor a protein in prokaryotic and preparation of monoclonal antibodies to PR-A

目的:制备孕激素受体A型单克隆抗体,为后期应用到免疫组化实验提供支持。方法:选择无缝克隆的方法将目的基因与载体连接到一起,通过诱导表达收集蛋白,经过免疫后细胞融合获得单克隆抗体。将该单克隆抗体经免疫组化验证。结果:将融合后筛选的阳性细胞制备获得的单克隆抗体7C7经免疫组化验证,发现其在乳腺癌、子宫肌瘤组织中呈现阳性,在直肠癌组织、平滑肌组织中呈现阴性,符合目的抗体的要求。结论:该方法获得的孕激素受体A型单克隆抗体较常规方法周期短,获得的抗体在不同组织间结果符合预判,对后期研究区别PR-A、PR-B,临床推广具有重要意义。

Objective： To prepare for m Ab of progesterone receptor. It would provide support for the immunohistochemistry behind. Methods： Target gene connected together with a carrier by seamless cloning method. The target protein that expression by inducing was collected. And with cell fusion method,the monoclonal antibodies were preparation. Then the m Ab were detected by IHC. Results： The m Ab（ clone 7C7） was detected and it found positive for the breast,uterine fibroid tissue,showed negative in colorectal cancer tissue,smooth muscle tissue,the goal of the claim were achieve. Conclusion： Finally,we found the method that prepare for m Ab was far beyond our imagination. The result of IHC on different samples about m Ab（ 7C7） obtained compliance with anticipation. Study on the difference between the PR-A and PR-B had significance.