Preparation and characterization of beta 1-bungarotoxin bispecific monoclonal antibody.

摘要

A hybrid hybridoma (tetradoma) that produces bispecific monoclonal antibodies (mAbs)2, which recognize two different epitopes on the A chain of beta 1-bungarotoxin (beta 1-bgt) at peptide sequences 46-51 and 100-106, has been obtained by fusing two hybridoma cell lines. The bispecific mAb were observed to inhibit 98% of the enzymatic activity of beta 1-bgt and neutralize its lethal toxicity completely. The avidity between the bispecific mAb and beta 1-bgt was noted to be 4.5 x 10(10) (litermol), which is about 45-150 folds higher than the avidity of its two parental mAbs. All the soluble complexes, obtained from bispecific mAb and beta 1-bgt with different molar ratios, emerged in the void volume of size exclusion chromatography column, indicating multiple complexes of beta 1-bgt and bispecific mAb were formed. Based on these results, it indicated that the binding of bispecific mAb with its two epitopes on beta 1-bgt, which facilitates the immuno-complex formation and enhances the avidity, also highly neutralizes the biological activity of beta 1-bgt.