PRODUCTION OF MONOCLONAL ANTIBODIES REACTIVE AGAINST CHOLINE ACETYLTRANSFERASE AND THEIR APPLICATION FOR STUDYING CHOLINERGIC SYSTEMS.

This paper has described experimental approaches for the development and application of monclonal antibodies against the specific cholinergic marker, choline acetyltransferase. Since ChAT was a difficult enzyme to purify, a strategy was developed for detecting the presence of specific antibodies when impure antigen preparations were used for immunization. The strategy included the use of a primary antigen binding assay, double-antibody co-precipitation, to measure antibody, and a radiochemical assay of enzyme activity to detect the presence of specific antigen. Hybridoma technology was then utilized to isolate and propagate clones of lymphoid cells producing antibodies of the desired specificity. Our experience exemplifies the advantages of monoclonal antibody approaches in that pure antigen preparations are not necessary as long as a suitably specific screening procedure is devised. Secondly, once desired clones are isolated, the availablity of continued antibody supplies is theoretically limitless.