分泌抗B型肉毒毒素单克隆抗体杂交瘤细胞系的建立

我国由肉毒毒素引起的食物中毒以A、B型最常见。但用常规法制备的抗血清在两型间常有交叉反应,不适于分型鉴定之用。即使应用亲和层析交叉吸收纯化,仍难以获得B型特异性抗体。为获得高效价的型特异性抗体,本研究以B型肉毒类毒素免疫纯系BALB/c小鼠,注射3次后,血清抗体琼脂双

HYBRIDOMA CELL LINES SECRETING MONOCLONAL ANTIBODIES AGAINST TYPE B TOXIN OF CLOSTRIDIUM BOTULINUM

Monoclonal antibodies against type B toxin of Clostridium Botulinum were prepared by fusing spleen cells, from BALB/c mouse immunized with the type B toxoid, with NS-.1 myeloma cell line. Screened by ELISA, 66.7% of hybridoma formed wells was spesific antibody positive. After four or five subcloning by limiting dilution, four different hybridoma cell lines (3B10, 3B11, 3G12, 4A5) were established and could secrete specific antibodies persistantly in culture medium as well as in BALB/c mice to produce antibody-enriched ascites. The results of testing the four monoclonal antibodies with type A and B toxoid showed that antibodies 3G12 and 4A5 were type B specific, but 3B10 and 3B11 had mild cross reaction with type A. Chromasoma assay has confirmed that the four cell lines are hybridomas.