A sensitive radioimmunoassay for adenosine in biological samples

✍ Scribed by Tomokazu Sato; Akira Kuninaka; Hiroshi Yoshino; Michio Ui

• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 806 KB
• Volume: 121
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90500-0

✦ Synopsis

A simple, sensitive, specific, and reproducible radioimmunoassay for the measurement of adenosine in biological materials has been developed. Adenosine antibody was obtained by immunizing rabbits with an immunogen prepared by conjugating 2',3'-disuccinyladenosine to human serum albumin. By succinylating adenosine in samples at the 2'-and 3'-0 positions with a premixed reagent consisting of succinic anhydride, triethylamine, and dioxane, the assay became sensitive enough to detect less than picomole amounts of adenosine in minute quantities of tissues. The cross-reactivity of structurally related compounds with the antibody was mostly negligible except for 2'-deoxyadenosine, whose usual concentration was very low. The use of this method made it possible to measure adenosine without any prior purification procedure. The immunoreactive materials in various biological samples disappeared during incubation of the samples with adenosine deaminase.

MATERIALS AND METHODS

Reagents

Sources of reagents were as follows: human serum albumin (crystallized), Miles Laboratories,