Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Mar;78(3):1901–1905. doi: 10.1073/pnas.78.3.1901

Enzyme-linked sandwich immunoassay for insulin using laser fluorimetric detection.

S D Lidofsky, W D Hinsberg 3rd, R N Zare
PMCID: PMC319243  PMID: 7015349

Abstract

Human serum samples are assayed for insulin by an enzyme-linked sandwich immunoassay. Horseradish peroxidase is used as an enzyme label for antibody, and enzyme activity is measured by means of the fluorogenic substrate, p-hydroxyphenylacetic acid. The product is detected by excitation of fluorescence with the 325-nm line of a continuous-wave helium/cadmium ion laser on line with reverse-phase high-pressure liquid chromatography. The incubation period is 90 min and the limit of detection of insulin is 30 pM, corresponding to 5 microunits/ml. This method correlates highly with radioimmunoassay, with coefficient of correlation r = 0.95.

Full text

PDF
1903

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arends J. Purification of peroxidase-conjugated antibody for enzyme immunoassay by affinity chromatography on concanavalin A. J Immunol Methods. 1979;25(2):171–175. doi: 10.1016/0022-1759(79)90052-8. [DOI] [PubMed] [Google Scholar]
  2. Cuatrecasas P. Insulin-sepharose: immunoreactivity and use in the purification of antibody. Biochem Biophys Res Commun. 1969 May 22;35(4):531–537. doi: 10.1016/0006-291x(69)90379-9. [DOI] [PubMed] [Google Scholar]
  3. Guilbault G. G., Brignac P. J., Jr, Juneau M. New substrates for the fluorometric determination of oxidative enzymes. Anal Chem. 1968 Jul;40(8):1256–1263. doi: 10.1021/ac60264a027. [DOI] [PubMed] [Google Scholar]
  4. Guilbault G. G., Sadar M. H., Peres K. Fluorometric determination of carbohydrates. Anal Biochem. 1969 Oct 1;31(1):91–101. doi: 10.1016/0003-2697(69)90245-0. [DOI] [PubMed] [Google Scholar]
  5. Kato K., Hamaguchi Y., Fukui H., Ishikawa E. Enzyme-linked immunoassay. I. Novel method for synthesis of the insulin-beta-D-galactosidase conjugate and its applicability for insulin assay. J Biochem. 1975 Jul;78(1):235–237. [PubMed] [Google Scholar]
  6. Kato K., Umeda Y., Suzuki F., Hayashi D., Kosaka A. Evaluation of a solid-phase enzyme immunoassay for insulin in human serum. Clin Chem. 1979 Jul;25(7):1306–1308. [PubMed] [Google Scholar]
  7. Morimoto Y., Yamamura Y. [Experimental studies on insulin enzyme-immunoassay kit, with special reference to the possibility of reducing the reaction time]. Horumon To Rinsho. 1978 Sep;26(9):987–989. [PubMed] [Google Scholar]
  8. Nose K. Purification and characterization of alkaline phosphatase from rat kidney. J Biochem. 1976 Feb;79(2):283–288. doi: 10.1093/oxfordjournals.jbchem.a131069. [DOI] [PubMed] [Google Scholar]
  9. Skelley D. S., Brown L. P., Besch P. K. Radioimmunoassay. Clin Chem. 1973 Feb;19(2):146–186. [PubMed] [Google Scholar]
  10. Wisdom G. B. Enzyme-immunoassay. Clin Chem. 1976 Aug;22(8):1243–1255. [PubMed] [Google Scholar]
  11. Yalow R. S. Radioimmunoassay: a probe for the fine structure of biologic systems. Science. 1978 Jun 16;200(4347):1236–1245. doi: 10.1126/science.208142. [DOI] [PubMed] [Google Scholar]
  12. Yoshioka M., Taniguchi H., Kawaguchi A., Kobayashi T., Murakami K., Seki M., Tsutou A., Tamagawa M., Minoda H., Baba S. Evaluation of a commercial enzyme immunoassay for insulin in human serum, and its clinical application. Clin Chem. 1979 Jan;25(1):35–38. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES