Chemiluminescent enzyme immunoassay for insulin

• Published in: BUNSEKI KAGAKU Vol. 36; no. 6; pp. 348 - 351
• Main Authors: AOYAGI, Shigeo, KUSUMI, Miyoko, MATSUYUKI, Akira, MAEDA, Masako, TSUJI, Akio
• Format: Journal Article
• Language: Japanese
• Published: The Japan Society for Analytical Chemistry 05.06.1987
• Subjects: addition of normal guinea pig serum; chemiluminescent enzyme immunoassay for insulin; determination of hydrogen peroxide; glucose oxidase; reduction of nonspecific binding
• ISSN: 0525-1931
• DOI: 10.2116/bunsekikagaku.36.6_348

The determination of hydrogen peroxide was performed by chemiluminescent and colorimetric methods.The light emitted from chemiluminescent reaction was measured with a Luminometer UPD-8000 (Meidensha Electric Mfg., Co., Ltd.). The detection limit was 1 pmol/tube in chemiluminescent method and 1 nmol/tube in colorimetric method. The calibration curves were obtained in the range from 1×10-8 to 1×10-3 M by chemiluminescent method and from 1×10-5 to 1×10-3 M by colorimetric method. In order to reduce nonspecific binding which reduces the sensitivity of sandwich enzyme immunoassay (EIA), the effect of addition of various proteins was studied. Guinea pig serum was found to be the most effective one. Basedon these experimental results, chemiluminescent EIA for insulin using β-D-glucose oxidase (GOD) as label enzyme was established. The anti insulin IgG-GOD was prepared by Ishikawa's method. The detection limit of this chemiluminescent EIA for insulin was 0.5 μU/tube and the regression line was Y (EIA)=1.21X(RIA) -3.60 (the correlation coefficient r=0.95).