As marijuana use has continued to grow, the need for accurate and reliable means of measuring cannabinoid levels has become more urgent. The objective of this Phase I project is to develop such a cannabinoid assay that will be rapid, non-isotopic, and capable of being performed in a field setting. Specific goals for Phase I will be to generate murine monoclonal antibodies to tetrahydrocannabinol (THC) and to begin work on the development of a sensitive and versatile THC fluorescence immunoassay that will be faster, easier, and at least as sensitive as currrent radioimmunoassays, which can detect 10 nanograms per milliliter.Monoclonal antibodies will be prepared against two different immunogens. A 51-carboxy 9-THC-BSA conjugate, which has been generously provided by NIDA, will first be used. Subsequently, a conjugate will also be used that will be prepared via carboxy-methylation and carbodiimide coupling of the phenolic hydroxyl of 9-THC. Approximately characterized antibodies will be attached to 0-galactosidase and used in a rapid solid-phase enzyme immunoassay using fluorogenic substrates. Such an assay system, when fully developed, will have significant commercial market, and will be useful for law enforcement agencies, the military, and other aspects of drug research programs.National Institute On Drug Abuse