孕激素受体A型原核表达及单克隆抗体制备

摘要

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7 C7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 mAb 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 mAb were detected by IHC. Results: The mAb ( 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 mAb was far beyond our imagination. The result of IHC on different samples about mAb(7C7)obtained compliance with an-ticipation. Study on the difference between the PR-A and PR-B had significance.