Selective clean-up method using an immunoaffinity column following radioimmunoassay of prostaglandin F2 alpha in biological fluids.
A selective clean-up method using an immunoaffinity column followed by radioimmunoassay (RIA) was developed for determining prostaglandin F2 alpha (PGF2 alpha) in human urine and plasma. Polyclonal antibody raised against PGF2 alpha, obtained from rabbits, was coupled to a tresyl-activated support based on a synthetic hydrophilic resin, TSKgel Tresyl-Toyopearl 650M, and used as the stationary phase for the immunoaffinity column. A human urine or plasma sample was introduced to this column, and PGF2 alpha was eluted with methanol-water (50:50, v/v) after the column had been washed. The eluate was subjected to competitive RIA for PGF2 alpha. The cross-reactivities of the RIA to a number of endogenous prostanoids, except PGD2, were negligible and the sensitivity was 4 pg/tube (p less than 0.05), giving a detection limit of 40 pg/ml when 1 ml of plasma or urine was available. The recoveries of plasma and urine samples were 98-108% and 96-106%, respectively, and their assay variances were 7-23%. The concentrations of endogenous PGF2 alpha in plasma and urine used here were estimated to be 72 and 98 pg/ml, respectively. This method should be very useful for various biological samples because of its good specificity, sensitivity, reliability and reproducibility.
Murai
Y
Y
Shionogi Research Laboratories, Shionogi & Co., Ltd., Osaka, Japan.
Ohno
M
M
Konishi
M
M
Kominami
G
G
D002846:Chromatography, Affinity; D003429:Cross Reactions; D015237:Dinoprost / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D007120:Immunochemistry; D007074:Immunoglobulin G / Q000276:immunology; D011453:Prostaglandins / Q000276:immunology; D011863:Radioimmunoassay; D012710:Serum Albumin, Bovine / Q000276:immunology
D007074:Immunoglobulin G; D011453:Prostaglandins; D012710:Serum Albumin, Bovine; D015237:Dinoprost