小鼠抗人亲环蛋白A(CypA)单克隆抗体的制备和应用  

Preparation and application of mouse monoclonal antibodies against human cyclophilin A( CypA)

作　　者：乔璐 洪欣[2] 李时孟 何成彦[1] 高申 王海[1] QIAO Lu;HONG Xin;LI Shimeng;HE Chengyan;GAO Shen;WANG Hai(Department of Laboratory Medicine,China-Japan Union Hospital of Jilin University,Changchun 130033,China;Vascular Surgery,China-Japan Union Hospital of Jilin University,Changchun 130033,China)

机构地区：[1]吉林大学中日联谊医院检验科,吉林长春130033 [2]吉林大学中日联谊医院血管外科,吉林长春130033

出　　处：《细胞与分子免疫学杂志》2018年第8期746-751,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81572082);吉林省科技厅资助项目(20140311092YY;20150101153;20150414015)

摘　　要：目的原核表达人亲环蛋白A (CypA)、制备抗人亲环蛋白A (CypA)单克隆抗体(mAb)并初步探索其在结肠癌发生发展中的作用。方法根据CypA基因序列,以原核表达系统表达CypA重组蛋白,经镍亲和层析柱纯化,免疫小鼠,杂交瘤法制备mAb;采用免疫组织化学染色法检测CypA在结直肠癌与癌旁组织中的差异表达,并分析与临床病理参数的相关性。结果成功构建了CypA原核表达载体,获得纯化的目的蛋白;获得1株稳定分泌抗人CypA mAb的杂交瘤细胞株;研究表明,结直肠癌组织CypA阳性染色率显著增高并与肿瘤分化程度及淋巴结转移相关。结论成功制备了小鼠抗人CypA mAb,并发现CypA在结直肠癌组织中高表达,与低分化和淋巴结转移呈正相关。Objective To prepare monoclonal antibodies against human cyclophilin A (CypA)and explore the role of CypA in the occurrence and development of colon cancer.Methods According to the CypA gene sequence,the recombinant protein of CypA was expressed by the prokaryotic expression vector and purified by a nickel affinity chromatography column. The monoclonal antibody was prepared by the antigen which is purified protein immunizing mice and hybridoma technique. Utilizing CypA monoclonal antibody,immunohistochemistry was used to detect the difference expression between colorectal cancer and adjacent tissues,meanwhile,analyse clinicopathological parameters.Results The prokaryotic expression vector of CypA was successfully constructed,so we obtain the purified protein;One hybridoma cell line secreting anti-CypA antibody was obtained.Immunohistochemical results showed that CypA expressed high level in colorectal cancer and relate with differentiation degree and lymph node metastasis.Conclusion The monoclonal antibody against human CypA was successfully produced in mice,and CypA was overexpressed in colorectal cancer tissues,which was positively correlated with low differentiation and lymph node metastasis.

关 键 词：亲环蛋白A 结直肠癌 单克隆抗体(mAb) 

分 类 号：Q813.2[生物学—生物工程] R392.11[医药卫生—免疫学]