A double-antibody enzyme immunoassay for determination of gonadotropin-releasing hormone in bovine plasma.

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  • GESHI Masaya
    Department of Animal Reproduction, College of Agriculture, University of Osaka Prefecture Tohoku National Agricultural Experiment Station
  • MATSUDA Kenji
    Department of Animal Reproduction, College of Agriculture, University of Osaka Prefecture
  • MORI Junichi
    Department of Animal Reproduction, College of Agriculture, University of Osaka Prefecture
  • KASUGA Naoki
    Research Division, Otsuka Assay Laboratory, Otsuka Pharmaceutical Co., Ltd.
  • IMAGAWA Kenichi
    Research Division, Otsuka Assay Laboratory, Otsuka Pharmaceutical Co., Ltd.
  • YANAIHARA Noboru
    University of Shizuoka, School of Pharmaceutical Sciences

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  • 牛における性腺刺激ホルモン放出ホルモンの二抗体法ｅｎｚｙｍｅ　ｉｍｍｕｎｏａｓｓａｙ
  • ウシ ニ オケル セイセン シゲキ ホルモン ホウシュツ ホルモン ノ 2 コ

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Abstract

A double-antibody enzyme immunoassay (EIA) for the determining GnRH in bovine plasma was studied. Antiserum raised against [Glu1]-GnRH-rabbit serum albumin conjugate in a male horse was used. Enzyme-labelled antigens were prepared by coupling synthetic GnRH with horseradish peroxidase using bis-diazotized benzidine. GnRH analogues and fragment of GnRH were tested for cross-reactivity in the EIA system. These results suggested that the C-terminus of GnRH molecule plays more important role in antigen-antibody binding than the N-terminus of it.<BR>Plasma specimens were extracted with acid-methanol. Process standard curve was suitable for plasma assay. The sensitivity of this EIA system was as little as 50 pg/tube of GnRH. After intravenous injection of 200 μg of synthetic GnRH in 2 steers, plasma concentrations of GnRH were determined in this EIA system. The levels abruptly increased just after injection and decreased thereafter.

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