THE OBJECTIVE OF THIS PROJECT IS TO USE IMMUNOLOGICAL METHODS TO PURIFY AND CHARACTERIZE THE 50 KDA ANOREXIGEN ISOLATED PREVIOUSLY IN LOW YIELDS USING CONVENTIONAL PROCEDURES.

THE OBJECTIVE OF THIS PROJECT IS TO USE IMMUNOLOGICAL METHODS TO PURIFY AND CHARACTERIZE THE 50 KDA ANOREXIGEN ISOLATED PREVIOUSLY IN LOW YIELDS USING CONVENTIONAL PROCEDURES. A SMALL AMOUNT OF THE PURIFIED ANOREXIGEN IS PRESENTLY AVAILABLE AND WILL BE USED TO GENERATE A SET OF MONOCLONAL ANTIBODIES. ONE OR MORE OF THESE ANTIBODIES WILL BE COUPLED TO AGAROSE WHICH WILL BE USED AS AN AFFINITY CHROMATOGRAPHY MATRIX FOR THE PURIFICATION OF THE 50 KDA ANOREXIGEN FROM RAT URINE. THIS METHOD OF PURIFICATION SHOULD BE A GREAT IMPROVEMENT OVER PREVIOUS METHODS AND SHOULD YIELD HOMOGENOUS ANOREXIGEN IN YIELDS AMPLE FOR THE PHYSIOLOGICAL AND BIOCHEMICAL STUDIES PROPOSED HERE. IT IS INTENDED THAT PHASE I WORK WILL LAY THE GROUNDWORK FOR THE PREPARATION OF PURE ANOREXIGEN. PHASE II WILL INVOLVE PURIFICATION OF SIGNIFICANT AMOUNTS OF THE PROTEIN, CHARACTERIZATION OF THE PROTEIN AND DEMONSTRATION OF SUSPECTED ACTIVITY OF POLYPEPTIDE FRAGMENTS. ULTIMATELY, RECOMBINANT DNA APPROACHES ARE ENVISIONED TO CLONE THE GENE FOR THE PROTEIN. IMMUNOLOGICAL AND GENE SEQUENCE CROSS HOMOLOGY WILL BE USED TO DETECT SIMILAR PROTEINS IN HUMANS. SUCH ANOREXIGEN MIGHT BE USEFUL IN CONTROLLING