HYBRIDOMA CELL LINES SECRETING MONOCLONAL ANTIBODIES AGAINST THE TYPE B TOXIN OF CLOSTRIDIUM BOTULINUM / 解放军医学杂志

ABSTRACT

The spleen cells from the BALB/C mouse immunized with the type B toxoid were fused with aNS-1 myeloma cell line. The superatant containing the hybridoma-formed cells with growing pores wasscreened by EL1SA, in which 66.7% the pores could secrete the specific antibodies against botulin.After a Subclonizing culture by the limiting dilution technique four hybridoma cell lines(3B10, 3B11, 3G12 and 4A5) were established and could secrete the specific antibodies persistently in the culture medium. in which antibody titers came to 10-3-10-5, while they were injected into the BALB/C mice intrape ritoneally ascites rich in antibodies with a titer of 10-5-10-8 was produced. The results testing the (our monoclonal antibodies with the type A and B toxoids showed that the antibodies of 3G12and 4A5 were specific for the type B toxoid, and those of 3B10 and 3B11 had light cross reaction with the type A toxoid. Identification of 1g showed that the antibodies of 3B10 and 3G12 were of IgG1, while those of 3B11 and 4A5 of IgG2. The chromosomal assay confirmed the four cell lines to be hybridoma. The neutroligation test in mice revealed that those four monoclonal antibodies did not show any protective effects on botulin.