Studies on establishment of methods for determination of growth hormone, insulin and thyroxine levels in serum of bovine by means of enzyme immunosorbentassay system

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In order to establish some methods for the determination of concentration of growth hormaone, insulin and thyroxine in the serum of bovine that be able to apply veterinary clinical medication in the bedside, modifications on the separation and purification methods of these hormones from anterior lobe of pituitary, ppancreas and thyroid gland and analytical conditions of enzyme immunosorbentassay (EIA) using avidin-biotin-peroxidase complexes (ABC) for the determination of bobine growth hormone (bGH), insulin and thyroxine (T_4) in serum of bovine were studied.1. bGH,insulin and T_4 was separated and purified from avobed mertioned tissues by well-known methods. We purified these further by HPLC and gel filtration chromatography. These protein fractions were recognized to be each a purified single fraction by SDS-PAGE and isoelectric focusing electriphoresis. Furthermore, those showed each a single band in the reaction with each purified antibodies by means of immunobotting. These protein fractions were isolated to be each of bGH,insulin and T_4 by their analytical techniques.2. The basical reactional conditions of EIA were studied using each extracted hormones, purified antibodies and ABC and alkaline phosphatase(ALP). We could t prepare the each standard curves, which showed stabilized reappearance.3. Concentrations of bGH,insulin and T_4 in serum from healthy and various patiant cattle were determined by means of two methods of EIA and dadioimmunoassay. Relatively significant correlation between these two tests was found, though data of EIA has shown a slightly high level more that RIA.4. Accordingly, it is considered necessary to modify about conjugation of antibody to solid phase and treatment of serum with some buffered solution for establishment of clinical EIA method.