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Chromatographic and radioimmunological methods for the determination of 5'-deoxy-5'-methylthioadenosine in biological
F Della Ragione1, A Oliva, V Gragnaniello
1Institute of Biochemistry of Macromolecules, I Medical School, University of Naples, Italy.
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.