Double antibody enzyme immunoassay for the quantitation of adenosine 3′,5′-cyclic monophosphate (Cyclic AMP) and guanosine 3′,5′-cyclic monophosphate (Cyclic GMP) in tissue and plasma

I. Yamamoto, J. Tsuji, T. Takai, M. Fujimoto

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A sensitive double antibody enzyme immunoassay for the quantitation of cyclic AMP and cyclic GMP is presented. Specific antisera to each nucleotide were raised in rabbits by immunization with succinyl cyclic nucleotide-human serum albumin conjugates. For competitive reaction, antibodies were incubated with a mixture of succinyl cyclic nucleotide labelled with $-D-galactosidase and unlabelled succinylated standard or sample cyclic nucleotides. The antibody-bound enzyme-hapten was separated from free hapten by anti-rabbit IgG immobilized to a polystyrene ball. Activity of the enzyme on the solid phase was fluorometrically determined. The assay system made it possible to ascertain values as low as 5 fmole of cyclic AMP or cyclic GMP. Cyclic nucleotides in plasma could be accurately determined by this method without requiring a deproteinizing reagent as the first step of assay. KEY WORDS: Enzyme immunoassay, Cyclic AMP, Cyclic GMP, $-D-galactosidase, Polystyrene ball. Activity of the enzyme on the solid phase was fluorometrically determined. The assay system made it possible to ascertain values as low as 5 fmole of cyclic ME' or cyclic GMP. Cyclic nucleotides in plasma could be accurately determined by this method without requiring a deproteinizing reagent as the first step of assay.

Original languageEnglish
Pages (from-to)173-196
Number of pages24
JournalJournal of Immunoassay
Volume3
Issue number2
DOIs
Publication statusPublished - 1982 Jan 1
Externally publishedYes

ASJC Scopus subject areas

  • Immunology
  • Pharmacology

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