免疫原性N6-异戊-2-烯基腺苷-蛋白质缀合物的新合成。相关抗体的制备、纯化及特异性。
New synthesis of immunogenic N6-isopent-2-enyladenosine-protein conjugate. Preparation, purification, and specificity of related antibodies.
作者信息
Papet M P, Delay D, Doumas P, Delmotte F
机构信息
Département de Biochimie des Glycoconjugués et Lectines Endogènes C.N.R.S., Orléans, France.
出版信息
Bioconjug Chem. 1992 Jan-Feb;3(1):14-9. doi: 10.1021/bc00013a002.
An original procedure which preserves the structure of the sugar ring is described to link a plant hormone as N6-isopent-2-enyladenosine [( 9R]iP) onto a protein carrier to prepare a more specific immunogen. This cytokinin is bound to bovine serum albumin (BSA) and ovalbumin by a five-step procedure. These [9R]iP-protein conjugates have a maximal absorption at 269 nm and show molar ratios of hormone bound to proteins in the range of 12:1 and 18:1 for BSA and ovalbumin, respectively. Polyclonal antibodies were raised in rabbits against [9R]iP-BSA and were purified by affinity chromatography. Titers and specificity of the antisera and purified antibodies were determined by ELISA and RIA. These antibodies are highly specific for [9R]iP and do not cross-react with zeatin and ribosylzeatin. An immunoaffinity matrix was prepared with a capacity of 1 microgram of [9R]iP/mL of gel.
本文描述了一种保留糖环结构的原始方法，即将植物激素N6 - 异戊 - 2 - 烯基腺苷[（9R）iP]连接到蛋白质载体上，以制备更具特异性的免疫原。这种细胞分裂素通过五步程序与牛血清白蛋白（BSA）和卵清蛋白结合。这些[9R]iP - 蛋白质缀合物在269 nm处有最大吸收，并且与BSA和卵清蛋白结合的激素摩尔比分别在12:1和18:1范围内。用[9R]iP - BSA在兔中制备多克隆抗体，并通过亲和层析进行纯化。通过ELISA和RIA测定抗血清和纯化抗体的效价和特异性。这些抗体对[9R]iP具有高度特异性，并且不与玉米素和核糖基玉米素发生交叉反应。制备了一种免疫亲和基质，其容量为每毫升凝胶1微克[9R]iP。
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