抗沙眼衣原体主要外膜蛋白(MOMP)单克隆抗体的制备及初步应用

Preparation and application of monoclonal antibodies against major outer membrane protein of Chlamydia trachomatis

制备抗沙眼衣原体(CT)单克隆抗体,建立沙眼衣原体的胶体金免疫层析快速检测方法。采用纯化沙眼衣原体主要外膜蛋白(MOMP)免疫Balb/c小鼠,选取高效价的免疫小鼠脾细胞与骨髓瘤细胞融合,ELISA检测筛选分泌抗MOMP单克隆抗体(McAb)的细胞株并进行相关鉴定。共获得分泌IgG1,IgG2b和IgM的3株单克隆抗体杂交瘤细胞株。纯化的单克隆抗体与鹦鹉热衣原体、肺炎衣原体、肺炎支原体均无交叉反应。通过位点分析选择配对良好的两株McAb,制备检测CT的双抗体夹心法免疫层析试纸条。制备的胶体金试纸条对MOMP的最低检出值可达50 ng·mL-1。

To prepare monoclonal antibodies （ McAb ） against Chlamydia trachomatis （ CT ） and establish the Chlamydia trachomatis GICA rapid detection method .Balb/c mice were injected with purified major outer membrane protein （ MOMP） of Chlamydia trachomatis.Spleen cells from immunized mice were fused with SP 2/0 myeloma cells.Hybirdoma cells which secrete antibodies against MOMP were selected and then identified .Three monoclonal antibodies which had very high affinity to MOMP were obtained .The isotypes of McAbs were IgG 1 , IgG2b and IgM. The above McAbs had no cross-reaction with Chlamydia psittaci, Chlamydia pneumoniae and Mycoplasma pneumoniae.Two McAbs with good pairing were selected to prepare GICA strip .The strip made up of these McAbs can be applied to detect CT .The sensitivity by GICA strip for detecting MOMP was about 50ng · mL^-1 .