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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
引用本文:连晓雯,杜惠芬,李克生. 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[J]. 浙江农业学报, 2014, 26(4): 872
作者姓名:连晓雯  杜惠芬  李克生
作者单位:甘肃省医学科学研究院,甘肃 兰州 730050
基金项目:沙眼衣原体抗原金标快速诊断试剂盒(项目编号:5CS0036-A52-031)
摘    要: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。

关 键 词:沙眼衣原体主要外膜蛋白  单克隆抗体  免疫层析试纸条  

Preparation and application of monoclonal antibodies against major outer membrane protein of Chlamydia trachomatis
LIAN Xiao wen,DU Hui fen,LI Ke sheng. Preparation and application of monoclonal antibodies against major outer membrane protein of Chlamydia trachomatis[J]. Acta Agriculturae Zhejiangensis, 2014, 26(4): 872
Authors:LIAN Xiao wen  DU Hui fen  LI Ke sheng
Affiliation:Gansu Medical Research Institute, Lanzhou 730050, china
Abstract: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 SP2/0 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 IgG1, 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 50 ng·mL-1.
Keywords:Chlamydia trachomatis major outer membrane protein  monoclonal antibody  munochromatographic test strip  
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