A simple and sensitive radioimmunoassay for adenosine.
We developed a simple and sensitive radioimmunoassay (RIA) for adenosine. The RIA is based on the double antibody method with adenosine 2', 3'-0-disuccinyl-3-[125I]-iodotyrosine methyl ester (diSc-adenosine-[125I]-TME) as a tracer. Anti-adenosine antiserum for the RIA was raised in rabbits immunized with diSc-adenosine conjugated to human serum albumin (diSc-adenosine-HSA). All samples and standards were succinylated prior to assay. The present immunoassay allows detection of 6.25-400 pmol/ml of adenosine in sample. Values obtained by the RIA and by a HPLC analysis showed a high correlation with correlation coefficient of 0.997. In order to determine adenosine in plasmas, blood cells must be separated in the presence of 6 mM EDTA, 0.006% dipyridamole (Dip) and 23 microM 2'-deoxycoformycin (dCF) at 2 degrees C. Adenosine in plasma could be accurately determined by the proposed method even without any pretreatments by deproteinizing. The adenosine levels with or without EDTA-treated normal human plasmas determined were 26.2 +/- 7.26 and 100 +/- 3.62 pmol/ml (mean +/- SEM), respectively.
Yamane
R
R
Immunology Laboratory, Yamasa Shoyu Co., Ltd., Chiba, Japan.
Nakamura
T
T
Matsuura
E
E
Ishige
H
H
Fujimoto
M
M
D000241:Adenosine / Q000031:analogs & derivatives / Q000097:blood* / Q000276:immunology; D000818:Animals; D001800:Blood Specimen Collection; D002851:Chromatography, High Pressure Liquid; D004176:Dipyridamole; D004285:Dogs / Q000097:blood; D004492:Edetic Acid; D006801:Humans; D008297:Male; D007470:Monoiodotyrosine / Q000031:analogs & derivatives / Q000276:immunology; D015649:Pentostatin; D011817:Rabbits; D011863:Radioimmunoassay; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C073983:adenosine 2',3'-O-disuccinyl-3-iodotyrosine methyl ester; D015649:Pentostatin; D004176:Dipyridamole; D004492:Edetic Acid; D007470:Monoiodotyrosine; D000241:Adenosine