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Radioimmunoassay of Prostaglandins F α , E 1 and E 2 in Human Plasma
Author(s) -
Dray F.,
Charbonnel B.,
Maclouf J.
Publication year - 1975
Publication title -
european journal of clinical investigation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.164
H-Index - 107
eISSN - 1365-2362
pISSN - 0014-2972
DOI - 10.1111/j.1365-2362.1975.tb02363.x
Subject(s) - radioimmunoassay , chemistry , luteal phase , chromatography , endocrinology , medicine , bovine serum albumin , carbodiimide , biochemistry , hormone , biology
. Antibodies against prostaglandins (PG)F 2α , E 1 and E 2 were obtained in rabbits immunized with respectively PG F 2α , PG E 1 and PG E 2 conjugated to bovine serum albumin by carbodiimide. A radioimmunoassay capable of measuring 7 pg of PG F α , 2 pg of PG E 2 and 14 pg of PG E 1 in human peripheral plasma is described. Plasma samples (pH 3, citric acid) are extracted with cyclohexane: ethyl acetate, 1:1 and then chromatographed on silicic acid columns to separate the prostaglandins into three fractions: fraction I, PG A, PG B and some unknown immunoreactive compounds; fraction II, PG E and fraction III, PG F α . The recovery is 80 % ± 6.2. Mean plasma levels in adults of PG F α and PG E, expressed in pg/ml: ‐PG F α 12 ± 2.8 (n = 25 men), 8 ± 2.3 (n = 18 women, follicular phase), 7 ± 1.4 (n= 18 women, luteal phase). ‐PG E l 40.5 ± 7.6 (n = 13 men), 38 ± 17.1 (n = 10 women). ‐PG E 2 4.5 ± 1 (n = 12 adult subjects). The major characteristics of the method described herein are the following: ‐ a large volume of plasma has to be processed (10 ml or more for PG Fa and PG E 1 , 5 ml or more for PG E 2 ). ‐ a chromatographic step is necessary to separate the different prostaglandins which makes it possible to circumvent problems of immunological cross reactivity and interference with unknown immunoreactive compounds. ‐ great care has been taken in collection of blood samples, especially to insure complete removal of blood cells namely platelets.