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.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2016年第7期1013-1016,共4页
Chinese Journal of Immunology
关键词
孕激素受体
原核表达
单克隆抗体
Progesterone receptor
Prokaryotic expression
Monoclonal antibody