Development of an enzyme-linked immunosorbent assay for 2,3-dinor-6-keto-prostagland in F1α in urine using a monoclonal antibody

✍ Scribed by Carmen Lindsay; Jean-Marie Moutquin; René C.-Gaudreault; Jean-Claude Forest

• Publisher: Elsevier Science
• Year: 1995
• Tongue: English
• Weight: 587 KB
• Volume: 28
• Category: Article
• ISSN: 0009-9120
• DOI: 10.1016/0009-9120(95)00018-5

✦ Synopsis

Objectives:

To develop and validate an enzyme-linked immunosorbent assay (elisa) for measurement of urinary 2,3-dinor-6-keto-prostaglandin f1 alpha (2,3d6kpgf1 alpha) 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 micrograms/l) used 2 mg/l of antibody and 3 micrograms/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 micrograms/l) of 2,3d6kpgf1 alpha added to an urinary sample was 101.2 +/- 6.3%. imprecision studies with non-pregnant (0.24 microgram/l) and pregnant (2.5 micrograms/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.