期刊论文

中文

现代免疫学

1001-2478

2015

35

5

393-398

30972693:国家自然科学基金 21473065:国家自然科学基金 2014CFA079:湖北省自然科学基金重点项目 CCNU13F011:中央高校基本科研业务费专项

本校为第一机构

研制抗人α-烯醇化酶的单克隆抗体,并对其生物学特性进行初步研究。利用原核表达系统对人α-烯醇化酶进行一系列重组表达。表达纯化后的人α-烯醇化酶作为抗原免疫BALB/c小鼠,采用杂交瘤技术以及克隆化培养筛选稳定分泌抗人α-烯醇化酶单克隆抗体的杂交瘤细胞株,采用免疫荧光技术、免疫组化技术对此单克隆抗体加以验证,并应用噬菌体随机十二肽库展示技术结合生物信息学方法对抗体识别表位进行初步鉴定。结果显示,克隆表达了人α-烯醇化酶,筛选获得一株稳定分泌抗人α-烯醇化酶单克隆抗体细胞株,此单克隆抗体的亚型为IgG1,它可与腹腔巨噬细胞的胞膜及胞浆的烯醇化酶特异性结合。噬菌体随机十二肽...

We set out to generate monoclonal antibodies against human α-enolase and identify the antigen-specific epitope. The recombinant human ccenolase protein was expressed by the prokaryotic system, and monoclonal antibodies against human α-enolase were generated. Epitopes of the mAb were analyzed by phage display peptide library. The specificity of the mAb was investigated by indirected ELISA, immunofluorescence （IF） and immunohistochemistry （IHC）. The results showed that human α-enolase was expressed successfully. A mAb against human α-enol...