ENZYME-IMMUNOASSAY FOR ALPHA-HUMAN ATRIAL NATRIURETIC POLYPEPTIDE - DIRECT MEASUREMENT OF PLASMA-LEVEL

被引:16
作者
HASHIDA, S
ISHIKAWA, E
NAKAO, K
MUKOYAMA, M
IMURA, H
机构
[1] MED COLL MIYAZAKI, DEPT BIOCHEM, MIYAZAKI 88916, JAPAN
[2] KYOTO UNIV, SCH MED, DEPT MED, DIV 2, KYOTO 606, JAPAN
[3] KYOTO UNIV, CTR RADIOISOTOPE RES, KYOTO 606, JAPAN
关键词
D O I
10.1016/0009-8981(88)90030-7
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
A sandwich enzyme immunoassay for .alpha.-human atrial natriuretic polypeptide (.alpha.-hANP) was developed. Polystyrene balls were coated with monoclonal IgG1 specific for the N-terminal half of the ring structure of .alpha.-hANP, and rabbit Fab'' specific for the C-terminal (17-28) of .alpha.-hANP was conjugated to horseradish peroxidase. The polystyrene ball was incubated with .alpha.-hANP standards or plasma and subsequently with the conjugate. Bound peroxidase activity was measured by fluorimetry. No addition of hANP-free plasma for the standard curve was required when <50 .mu.l of plasma sample was used. The detection limit of .alpha.-hANP was 30 fg (10 amol). Using 50 .mu.l of plasma, we could detect 0.6 ng (0.2 pmol)/l without extraction. The basal plasma .alpha.-hANP level of healthy men in a supine position after an overnight fast was 24.5 .+-. 13.2 (SD) ng/l and tended to decrease (15.3 .+-. 8.5 (SD) ng/l) after intravenous administration of furosemide (40 mg) and subsequent one hour walking. There was a highly significant correlation between the plasma .alpha.-hANP concentrations measured by this enzyme immunoassay and by radioimmunoassay (r=0.92). This enzyme immunoassay for .alpha.-hANP allows measurement of plasma .alpha.-hANP without extraction even in the volume-contracted state.
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页码:11 / 18
页数:8
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