用于测定生物体液中5'-脱氧-5'-甲硫基腺苷的色谱法和放射免疫法。

Chromatographic and radioimmunological methods for the determination of 5'-deoxy-5'-methylthioadenosine in biological fluids.

作者信息

Della Ragione F, Oliva A, Gragnaniello V, Fioretti M, Fioretti A, Menna L F, Papparella V, Zappia V

机构信息

Institute of Biochemistry of Macromolecules, I Medical School, University of Naples, Italy.

出版信息

J Chromatogr. 1988 May 25;440:141-9. doi: 10.1016/s0021-9673(00)94518-7.

DOI:10.1016/s0021-9673(00)94518-7

PMID:3403660

Abstract

Two specific methods for the determination of 5'-deoxy-5'-methylthioadenosine (MTA) in biological samples have been developed. The chromatographic procedure requires a preliminary step on a phenylboronate column to remove non-cis-diol compounds. The sample is then analysed using a high-performance liquid chromatography system equipped with a reversed-phase column. 5'-Deoxy-5'-methyl-thio[2-3H]adenosine with high specific activity was synthesized and employed as an internal standard. An alternative radioimmunoassay (RIA) procedure has also been developed. The RIA method is based on competition between the unlabelled thio-ether and 3H-labelled MTA for the binding to a specific antiserum. Anti-MTA antibodies were obtained from rabbits immunized with the nucleoside covalently linked to carrier proteins. Both the chromatographic and RIA procedures gave identical results when employed to determine MTA in human urine.

摘要

已开发出两种测定生物样品中5'-脱氧-5'-甲硫基腺苷（MTA）的具体方法。色谱法需要在苯基硼酸酯柱上进行预处理以去除非顺式二醇化合物。然后使用配备反相柱的高效液相色谱系统对样品进行分析。合成了具有高比活性的5'-脱氧-5'-甲硫基[2-³H]腺苷并用作内标。还开发了一种替代的放射免疫测定（RIA）方法。RIA方法基于未标记的硫醚和³H标记的MTA之间竞争与特异性抗血清结合。抗MTA抗体是从用与载体蛋白共价连接的核苷免疫的兔子中获得的。当用于测定人尿中的MTA时，色谱法和RIA方法都给出了相同的结果。