The long term goal of the project is to study the feasibility of using human monoclonal antibodies as detoxification agents against drugs of abuse and to study the molecular biology of drug receptors in the brain.
The specific aims of the present proposal are focused on the production and testing of human anti- phencyclidine monocolonal antibodies and on the design of a biochemical marker of phencyclidine receptors in the brain. Phencyclidine (1-( 1-phenylcyclohexy ) piperidine, PCP, angel dust) is among the most commonly available of the abused drugs; an antidote or detoxification agent is needed to treat phencyclidine toxicity. A biochemical marker is needed to further advance the molecular biological study of the PCP receptor. Plans are made to produce a library of anti- phencyclidine hybridomas, to characterize the immunological and biochemical properties of the monoclonal antibodies, to examine the detoxification effects of these antibodies in biochemical models, and to produce an anti-idiotype monoclonal antibody to be used as a high affinity antagonist of PCP receptors in the central nervous system. Anti-PCP hybridomas will be produced by using the mouse- human heteromyeloma as fusion partners for the in vitro sensitized human B cell populations. A library of human monoclonal hybridomas secreting anti-PCP antibodies will be cultured and studied. A large number of these monoclonal antibodies needs to be characterized in order to select the optimal antibody for use as a detoxification agent, and as immunogens to produce anti-idiotype hybridomas. The immunological specificity and equilibrium binding constant of these monoclonal antibodies will be tested against a panel of phencyclidine-related compounds. The ability of the anti-PCP immunoglobulin (or FAB) in reducing the effective PCP concentration in in vitro systems will also be studied. Acetylcholine receptor serves as an ideal model receptor system for biochemical studies. The ability of the monoclonal antibodies in reversing the binding of PCP to the acetylcholine receptor will be tested. The anti-phencyclidine antibodies will also be used as immunogens in the mouse to produce mouse anti-idiotype hybridomas. High affinity anti-idiotype antibodies will be selected as possible antagonists for studying the PCP receptor in the brain.
