Determination of prostacyclin in plasma through a bioluminescent immunoassay for 6-keto-prostaglandin F: Implication of dosage in patients with primary pulmonary hypertension

Urvee A. Desai, Sapna K. Deo, Kenneth V. Hyland, Michael Poon, Sylvia Daunert

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

This work describes a solid-phase immunoassay for 6-keto-prostaglandin F, the stable hydrolysis product of prostacyclin (prostaglandin I2). Prostacyclin, a potent vasodilator with antiplatelet and antiproliferative properties is an effective treatment for primary pulmonary hypertension and pulmonary arterial hypertension associated with scleroderma and scleroderma-like syndrome. Levels of 6-keto-prostaglandin F can be directly correlated with levels of prostacyclin. Therefore, 6-ketoprostaglandin F has become the indicator of choice to measure prostacyclin levels. The single-step immunoassay for 6-keto-prostaglandin F reported here was developed using the bioluminescent protein aequorin as a label. Analyte-label conjugates were constructed by linking the carboxyl group of 6-keto-prostaglandin F and lysine residues of aequorin by chemical conjugation methods. The binding properties of 6-keto-prostaglandin F toward its antibody and the bioluminescent properties of aequorin were retained in the conjugate, which was then used to generate a dose-response curve for the analyte in a convenient microtiter plate format. The concentration of 6-keto-prostaglandin F after extraction from plasma showed good correlation with the concentration of 6-ketoprostaglandin F obtained without prior extraction of the same plasma sample. This measurement demonstrated that the assay allows the measurement of 6-keto-prostaglandin F directly in plasma without any pretreatment of the samples, which results in a much simpler method with a faster assay time.

Original languageEnglish (US)
Pages (from-to)3892-3898
Number of pages7
JournalAnalytical Chemistry
Volume74
Issue number15
DOIs
StatePublished - Aug 1 2002
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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