Singh V
Institute of Self-Organizing Systems and Biophysics, North-Eastern Hill University, Meghalaya, India.
Biochem Int. 1991 Oct;25(3):509-20.
In order to render hypothalamic 'self' decapeptide GnRH immunogenic, the GnRH and its azotized derivative were covalently coupled to the carrier protein, BSA. The azotization at histidine and or tyrosine of GnRH was carried out and characterized extensively. Three different molar ratios of GnRH/azo-GnRH with BSA were prepared and characterized by physico-chemical techniques. The conjugates purified by gel-filtration chromatography on G-100 and on gel permeation column using HPLC were subjected to molar ratio determination by amino acid analysis. The immunoreactivity of GnRH/azo-GnRH towards monoclonal and polyclonal antibodies was drastically hampered after conjugation. It is therefore suggested that in the GnRH-carrier conjugate, GnRH concentration should be determined carefully by using specific radioimmunoassay.
为使下丘脑“自身”十肽促性腺激素释放激素（GnRH）具有免疫原性，将GnRH及其偶氮化衍生物与载体蛋白牛血清白蛋白（BSA）共价偶联。对GnRH的组氨酸和/或酪氨酸进行偶氮化，并进行了广泛的表征。制备了GnRH/偶氮-GnRH与BSA的三种不同摩尔比，并通过物理化学技术进行了表征。通过在G-100上进行凝胶过滤色谱以及使用高效液相色谱（HPLC）在凝胶渗透柱上纯化的偶联物，通过氨基酸分析进行摩尔比测定。偶联后，GnRH/偶氮-GnRH对单克隆抗体和多克隆抗体的免疫反应性受到严重阻碍。因此建议，在GnRH-载体偶联物中，应使用特异性放射免疫测定法仔细测定GnRH的浓度。
