Interaction of monoclonal antibodies with mammalian choline acetyltransferase

摘要: Abstract Monoclonal antibodies selective for rat brain choline acetyltransferase (acetyl-CoA: O-acetyltransferase, EC 2.3.1.6) were prepared by standard techniques. Five cell lines isolated from spleen cell-SP/2 hybrids repetitive cloning with a screening method that used the intrinsic activity of enzyme. All secrete immunoglobulin mouse subclass IgGl, and none inhibit enzyme directly. The size antibody-enzyme immune complexes formed different pairs monoclonal was determined gel filtration HPLC. By comparing elution position acetyl-transferase after incubation paired antibodies, spatial relationship antibody binding domains relative to each other can be defined classified as independent, mutually exclusive, or overlapping. Immune in excess Mr 600,000 some antigen, indicating independent on In one case, an complex only 300,000, they bound exclusive fashion. most cases, reacted give simultaneously both higher lower complexes. We conclude all five bind relatively localized region surface. Antibodies screened usefulness immunohistochemical markers acetyltransferase-containing neurons using indirect immunoperoxidase method. One intensely stains bodies motor processes manner spinal cord stem aldehyde-fixed tissue; remaining do not react fixed tissue.