Abstract
This work describes a solid-phase immunoassay for 6-keto-prostaglandin F1α, 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 F1α can be directly correlated with levels of prostacyclin. Therefore, 6-ketoprostaglandin F1α has become the indicator of choice to measure prostacyclin levels. The single-step immunoassay for 6-keto-prostaglandin F1α 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 F1α and lysine residues of aequorin by chemical conjugation methods. The binding properties of 6-keto-prostaglandin F1α 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 F1α after extraction from plasma showed good correlation with the concentration of 6-ketoprostaglandin F1α obtained without prior extraction of the same plasma sample. This measurement demonstrated that the assay allows the measurement of 6-keto-prostaglandin F1α directly in plasma without any pretreatment of the samples, which results in a much simpler method with a faster assay time.
Original language | English (US) |
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Pages (from-to) | 3892-3898 |
Number of pages | 7 |
Journal | Analytical Chemistry |
Volume | 74 |
Issue number | 15 |
DOIs | |
State | Published - Aug 1 2002 |
Externally published | Yes |
ASJC Scopus subject areas
- Analytical Chemistry