期刊论文

Wu, Y.-M.

中文

中华微生物学和免疫学杂志

0254-5101

2011

31

11

1041-1045

10.3760/cma.j.issn.0254-5101.2011.11.019

国家自然科学基金（30970165、81102230） 湖南省自然科学基金（09JJ3059） 湖南省高校科技创新团队资助项目（湘教通[20103212号）

本校为第一且通讯机构

目的纯化抗沙眼衣原体pORF5单克隆抗体2H4并鉴定其免疫学特性。方法大量培养pORF5单克隆抗体阳性杂交瘤细胞株并收集培养上清，采用G蛋白免疫亲和层析法纯化2H4单克隆抗体；酶联免疫吸附试验（ELISA）测定2H4效价及抗体亚类；Western blot鉴定其特异性；免疫荧光试验（IFA）检测2H4单克隆抗体的衣原体种属特异性。结果纯化后2H4抗体的纯度高达93%；效价为1∶1024，免疫球蛋白类型为IgG2a；2H4抗体不仅能特异性识别pORF5融合蛋白，而且能特异性识别Ct血清型A、D、L2、鼠衣原体（MoPn）、鹦鹉热嗜衣原体（6BC）质粒所编码的内源性pORF5蛋白，但不识别衣原体其他质粒蛋白和肺炎嗜衣原体（Cpn）。结...

Objective: To purify and characterize the monoclonal antibody (McAb) against Chlamydia trachomatis pORF5 plasmid protein. Methods: The hybridoma cells stably secreting specific McAb against pORF5 were cultured in a large scale, and protein G purification by affinity chromatography was used to purify 2H4 McAb. ELTSA was used to determine the antibody titer, and identify McAb isotype. Immunofluorescence assay (IFA) and Western blot were performed to detect McAb specificity. Results: The purity of 2H4 antibody was 93%, the titer reached 1:1024, and 2H4 McAb was identified to belong to TgG2a isoty...