抗麻风杆菌酚糖脂1末端三糖的单克隆抗体的制备

Generation of Monoclonal Antibodies Against Terminal Trisaccharide of Phenolic Glycolipid I of Mycobacterium Leprae

将麻风杆菌特异性抗原酚糖脂Ⅰ(phenolic glycolipid I,PGL-I)包被在明尼苏达沙门氏菌(Salmonella minisota)表面,制成免疫原、脾脏内免疫BALB/c鼠,取免疫鼠脾脏制成脾细胞悬液,与Sp2/0细胞按常法融合,选择性培养。凡有克隆生长的,对其上清,用人工合成、含PGL-I末端三糖的人工半合成抗原——NT-O-BSA作ELISA进行筛选。从中选择了一株对PGL-I末端三糖表位特异的单克隆抗体——E_(10)F_1进行了初步鉴定。E_(10)F_1的Ig亚类为IgG3,与大多数抗糖抗体的Ig亚类一致。经ELISA及斑点免疫吸附试验鉴定,E_(10)F_1仅与PGL-I包被的明尼苏达沙门氏菌出现阳性反应,不与明尼苏达沙门氏菌、BSA、BCG、耻垢分枝杆菌和堪萨斯分枝杆菌发生反应。证明单克隆抗体E_(10)F_1所识别的抗原表位为PGL-I分子末端三糖,也是麻风杆菌的特异性抗原表位。文中对制备抗糖的单克隆抗体的实验设计进行了讨论。

BALB/c mice were immunized with phenolic glycolipid Ⅰ of mycobacterium leprae (PGL-Ⅰ) that coated on salmonella minisota, by intrasplenic injection. The immuned mouse spleen cells were fused with Sp2/0 myeloma cells. Hybridomas were screened by artificial NT-O-BSA, which bears the specific epitope-trisaccha-ride of PGL-I. Monoclonal antibody specific for the terminal trisacchar ide of PGL- I (E_(10)F_1) was derived. This conclusion was bassed on the following observation:E_(10)-F_1 can bind to NT-O-BSA and salmonella minisota coated with PGL-Ⅰ, but it can't react with mycobacterium smegmatis, mycobacterium kansas, BCG and salmonella minisota without coating.The experimental design for development of a monoclonal antibody against carbohydrate has been discussed.