Preparation and characterization of beta 1-bungarotoxin bispecific monoclonal antibody.
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) (liter/nmol), 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.
Yang
C C
CC
Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan, Republic of China.
Chan
H L
HL
D018033:Antibodies, Bispecific / Q000096:biosynthesis*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex / Q000276:immunology; D002038:Bungarotoxins / Q000737:chemistry / Q000276:immunology*; D002459:Cell Fusion; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D006825:Hybridomas; D007140:Immunoglobulin Fab Fragments / Q000276:immunology
D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002038:Bungarotoxins; D004791:Enzyme Inhibitors; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments