Abstract
We developed a simple and sensitive radioimmunoassay (RIA) for adenosine. The RIA is based on the double antibody method with adenosine 2',3'-O-disuccinyl-3-[125I]-iodotyrosine methyl ester (diSc-adenosine-[123I]-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 μM 2’-deoxycoformycin (dCF) at 2°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. (KEY WORDS: adenosine, radioimmunoassay (RIA), dipyridamole, plasma adenosine level, EDTA, deoxycoformycin).
| Original language | English |
|---|---|
| Pages (from-to) | 501-519 |
| Number of pages | 19 |
| Journal | Journal of Immunoassay |
| Volume | 12 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Dec 1 1991 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Immunology
- Pharmacology
Cite this
A simple and sensitive radioimmunoassay for adenosine. / Yamane, Ryota; Nakamura, Takeshi; Matsuura, Eiji; Ishige, Hideyuki; Laboratory, Immunology; Division, Diagnostics; Fujimoto, Masao.
In: Journal of Immunoassay, Vol. 12, No. 4, 01.12.1991, p. 501-519.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A simple and sensitive radioimmunoassay for adenosine
AU - Yamane, Ryota
AU - Nakamura, Takeshi
AU - Matsuura, Eiji
AU - Ishige, Hideyuki
AU - Laboratory, Immunology
AU - Division, Diagnostics
AU - Fujimoto, Masao
PY - 1991/12/1
Y1 - 1991/12/1
N2 - We developed a simple and sensitive radioimmunoassay (RIA) for adenosine. The RIA is based on the double antibody method with adenosine 2',3'-O-disuccinyl-3-[125I]-iodotyrosine methyl ester (diSc-adenosine-[123I]-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 μM 2’-deoxycoformycin (dCF) at 2°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. (KEY WORDS: adenosine, radioimmunoassay (RIA), dipyridamole, plasma adenosine level, EDTA, deoxycoformycin).
AB - We developed a simple and sensitive radioimmunoassay (RIA) for adenosine. The RIA is based on the double antibody method with adenosine 2',3'-O-disuccinyl-3-[125I]-iodotyrosine methyl ester (diSc-adenosine-[123I]-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 μM 2’-deoxycoformycin (dCF) at 2°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. (KEY WORDS: adenosine, radioimmunoassay (RIA), dipyridamole, plasma adenosine level, EDTA, deoxycoformycin).
UR - http://www.scopus.com/inward/record.url?scp=0025837332&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0025837332&partnerID=8YFLogxK
U2 - 10.1080/01971529108053277
DO - 10.1080/01971529108053277
M3 - Article
C2 - 1806586
AN - SCOPUS:0025837332
VL - 12
SP - 501
EP - 519
JO - Journal of Immunoassay and Immunochemistry
JF - Journal of Immunoassay and Immunochemistry
SN - 1532-1819
IS - 4
ER -