In vitro neutralization by monoclonal antibodies of alpha-bungarotoxin binding to acetylcholine receptor.
In order to develop monoclonal antibodies that would neutralize binding of alpha-bungarotoxin to acetylcholine receptor in vitro, mice were hyperimmunized with native toxin. Frequent small doses of toxin were used. Hybridoma supernatants were screened by ELISA and six monoclonal antibodies isolated and tested. The anti-alpha-bungarotoxin monoclonal antibodies consisted of IgM, IgG1 or IgG2a antibodies. In an in vitro neutralization assay measuring the effect of the antibodies on the binding of iodinated alpha-bungarotoxin to BC3H1 and TE671 (mouse and human cell lines bearing acetylcholine receptor), three of the six monoclonal antibodies were able to neutralize toxin binding. These studies demonstrate the feasibility of using native toxin for the generation of hybridomas, and the potential of using in vitro neutralization assays to screen hybridomas for in vivo neutralization.
Pachner
A R
AR
Department of Neurology, Georgetown University Hospital, Washington, DC 20007.
Ricalton
N
N
D000818:Animals; D000911:Antibodies, Monoclonal; D002038:Bungarotoxins / Q000276:immunology / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D009500:Neutralization Tests; D011950:Receptors, Cholinergic / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; D011950:Receptors, Cholinergic