Enzyme immunoassays for the estimation of adenosine 3′,5′ cyclic monophosphate and guanosine 3′,5′ cyclic monophosphate in biological fluids

J. K. Horton, R. C. Martin, S. Kalinka, A. Cushing, J. P. Kitcher, M. J. O'Sullivan, P. M. Baxendale

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Simple, rapid and sensitive competitive enzyme immunoassays for the estimation of adenosine 3′,5′ cyclic monophosphate (cAMP) and guanosine 3′,5′ cyclic monophosphate (cGMP) in human plasma and urine are described. Specific antisera to each nucleotide were raised in rabbits by immunization with succinyl cyclic nucleotide - human serum albumin conjugates. For the assay, specific antibodies were incubated with a mixture of succinyl cyclic nucleotide labelled with horseradish peroxidase together with unlabelled standard or sample. The antibody-bound enzyme conjugate was separated from free hapten by anti-rabbit (IgG) sera immobilized to a microtitre plate. Activity of the bound enzyme conjugate was determined with tetramethylbenzidine. The assays were capable of detecting levels as low as 2 fmol of cAMP and cGMP. Good correlations were obtained between values generated by enzyme immunoassay and radioimmunoassay.

Original languageEnglish
Pages (from-to)31-40
Number of pages10
JournalJournal of Immunological Methods
Volume155
Issue number1
DOIs
StatePublished - Oct 19 1992
Externally publishedYes

Fingerprint

Guanosine Monophosphate
Cyclic Nucleotides
Cyclic GMP
Immunoenzyme Techniques
Adenosine
Rabbits
Antibodies
Haptens
Guanosine
Enzymes
Horseradish Peroxidase
Serum Albumin
Radioimmunoassay
Immune Sera
Immunization
Nucleotides
Urine
Serum
anti-IgG

Keywords

  • Adenosine 3′,5′ cyclic monophosphate
  • Enzyme immunoassay
  • Guanosine 3′,5′ cyclic monophosphate

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Enzyme immunoassays for the estimation of adenosine 3′,5′ cyclic monophosphate and guanosine 3′,5′ cyclic monophosphate in biological fluids. / Horton, J. K.; Martin, R. C.; Kalinka, S.; Cushing, A.; Kitcher, J. P.; O'Sullivan, M. J.; Baxendale, P. M.

In: Journal of Immunological Methods, Vol. 155, No. 1, 19.10.1992, p. 31-40.

Research output: Contribution to journalArticle

Horton, J. K. ; Martin, R. C. ; Kalinka, S. ; Cushing, A. ; Kitcher, J. P. ; O'Sullivan, M. J. ; Baxendale, P. M. / Enzyme immunoassays for the estimation of adenosine 3′,5′ cyclic monophosphate and guanosine 3′,5′ cyclic monophosphate in biological fluids. In: Journal of Immunological Methods. 1992 ; Vol. 155, No. 1. pp. 31-40.
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abstract = "Simple, rapid and sensitive competitive enzyme immunoassays for the estimation of adenosine 3′,5′ cyclic monophosphate (cAMP) and guanosine 3′,5′ cyclic monophosphate (cGMP) in human plasma and urine are described. Specific antisera to each nucleotide were raised in rabbits by immunization with succinyl cyclic nucleotide - human serum albumin conjugates. For the assay, specific antibodies were incubated with a mixture of succinyl cyclic nucleotide labelled with horseradish peroxidase together with unlabelled standard or sample. The antibody-bound enzyme conjugate was separated from free hapten by anti-rabbit (IgG) sera immobilized to a microtitre plate. Activity of the bound enzyme conjugate was determined with tetramethylbenzidine. The assays were capable of detecting levels as low as 2 fmol of cAMP and cGMP. Good correlations were obtained between values generated by enzyme immunoassay and radioimmunoassay.",
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AU - Cushing, A.

AU - Kitcher, J. P.

AU - O'Sullivan, M. J.

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