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Title of article :
Development of an enzyme-linked immunosorbent assay for 2,3-dinor-6-keto-prostagland in F1α in urine using a monoclonal antibody
Author/Authors :
Carmen Lindsay، نويسنده , , Jean-Marie Moutquin، نويسنده , , René C.-Gaudreault، نويسنده , , Jean-Claude Forest، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 1995
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Abstract :
Objectives: To develop and validate an enzyme-linked immunosorbent assay (ELISA) for measurement of urinary 2,3-dinor-6-keto-prostaglandin F1α (2,3D6KPGF1α) using a monoclonal antibody and a horseradish peroxidase-linked antigen. Design and Methods: Assay validation included optimization of the standard curve, antibody cross-reactivity, accuracy and imprecision studies together with preliminary measurement of clinical samples. Results: Optimal conditions of the standard curve (0.078–10.0 μg/L) used 2 mg/L of antibody, and 3 μg/L of peroxidase conjugate in each well, at pH 7.2. The coefficient of variation of various concentrations of the standard curve averaged 6.8%. Antibody cross-reactivity was <0.01% for related prostanoids. Recovery of known amounts (0.1–5.0 μg/L) of 2,3D6KPGF1α added to an urinary sample was 101.2 ± 6.3%. Imprecision studies with non-pregnant (0.24 μg/L) and pregnant (2.5 μg/L) samples displayed an intraassay variability of 8.9 and 9.9%, and an interassay variability of 9.6 and 10.0%, respectively. Urinary measurements in the non-pregnant and pregnant states were similar to those previously reported. An apparent decreased concentration was observed early in pregnancy in future preeclampsia. Conclusion: With similar precision and validity, our assay method is time- and cost-saving. Preliminary urinary measurements show that this analyte may be of interest as an early marker for preeclampsia.
Keywords :
2 , ELISA , hypertension , monoclonal antibody , Preeclampsia , 3-dinor-6-keto-prostaglandin F 1 , urinary prostacyclin metabolite
Journal title :
Clinical Biochemistry
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