用酶联免疫吸附受体法检测鱼类生长激素的生物活性

ELISA-RECEPTOR ASSAY FOR TESTING THEBIOACTIVITY OF FISH GROWTH HORMONE

根据激素—受体反应的原理，结合酶联免疫吸附测定法（ＥＬＩＳＡ）和放射受体测定法（ＲＲＡ）的优点，应用我们纯化的草鱼生长激素（ｇｃＧＨ）和大鳞大马哈鱼生长激素（ｓＧＨ）及其特异抗体．采用鱼类肝细胞膜受体制剂，首次建立了测定鱼类ＧＨ生物活性的酶联免疫吸附受体测定法（ＥＬＩＳＡ—ＲＡ）。此法检测草鱼ＧＨ的灵敏度达０．０６３～０．１２５μｇ／ｍｌ，检测大马哈鱼ＧＨ与草鱼肝膜受体结合的灵敏度达０．２５μｇ／ｍｌ。同时表明草鱼ＧＨ、基因重组鲤ＧＨ以及大马哈鱼ＧＨ均可不同程度地与草鱼（Ｃｔｅｎｏｐｈａｒｙｎｇｏｄｏｎｉｄｅｌｌｕｓ）、鲤（Ｃｙｐｒｉｎｕｓｃａｒｐｉｏ）、鲶（Ｓｉｌｕｒｕｓｍｅｒｉｄｉｏｎａｔｉｓ）和黄颡鱼（Ｐｅｔｔｅｏｂａｇｒｕｓｆｕｌｖｉｄｒａｃｏ）肝细胞ＧＨ受体特异结合。本测定系统可用于测定鱼类垂体中的ＧＨ含量以及监测鱼类ＧＨ分离纯化过程。

According to the principle of hormone-receptor specific binding and combin ing the advantages of the enzyme-linked immunosorbent assay (ELISA ) and radioreceptorassay (RRA ),we developed an ELISA-receptor assay (ELISA-RA ) for testing the bioactivi ty of fish growth hormone. In this ELISA-RA system, the fish GH molecules can bind toboth fish liver membrane receptors and its antibodies. This method can examine both bioac tivity and immunorcactivity of fish GH. In the case of grass carp liver membrane as receptor,the sensitivity of this assay is 0. 063-0. 125μg/ml for grass carp GH and0. 25μg/ml for chinook salmon (Oncorhy nchus tschawytscha ) GH, respectively. The presentstudy also shows that grass carp GH, recombinant common carp GH and chinook salmon GHcan specifically bind to liver membranes of common carp (Cyprinus carpio), southern sheat fish (Silurs meridionalis), yellow catfish (Pelteobagrus fulvidraco ) as well as grass carp(Ctenop haryng od on of ellus).