Development of a Microtiter Plate Enzyme-linked Immunosorbent Assay for 17 α, 20 β-21-trihydroxy-4-pregnen-3-one, a Teleost Gondadal Steroid

A simple solid-phase microtiter plate enzyme-linked immunosorbent assay (ELISA) for 17ƒ¿ , 20ƒÀ, 21-trihydroxy-4-pregnen-3-one (20ƒÀ-S), a teleost maturation inducing steroid, was developed using 20ƒÀ-S-3-CMO-horseradish peroxidase as the label, and a homologous antiserum raised in rabbits. Second antibody coated to the plate was used to capture the antiserum with labeled or unlabeled 20/3-S. Within three hours, a steep standard curve covering 1.5-192 ƒÊg/well was obtained. The sensitivity and precision of this ELISA was almost the same as those of ordinary steroid radioimmunoassays. This non-isotopic assay system also allows prompt measurement of 20ƒÀ-S in catfish plasma with good specificity.

[1]  D. Kime,et al.  In vitro biosynthesis of steroids, including 11-deoxycortisol and 5α-pregnane-3β,7α,17,20β-tetrol, by ovaries of the goldfish Carassius auratus during the stage of oocyte final maturation , 1992 .

[2]  K. Aida,et al.  Plasma Steroid Hormone Profiles during HCG Induced Ovulation in Female Walking Catfish Clarias batrachus(Endocrinology) , 1992 .

[3]  P. Thomas,et al.  Isolation of a novel maturation-inducing steroid produced in vitro by ovaries of Atlantic croaker. , 1989, General and comparative endocrinology.

[4]  G. J. Marcus,et al.  Estradiol assay by microtitre plate enzyme immunoassay. , 1988, Journal of steroid biochemistry.

[5]  H. M. Dye,et al.  The effects of stress on plasma ACTH, α-MSH, and cortisol levels in salmonid fishes , 1986 .

[6]  G. J. Marcus,et al.  A simple enzyme-linked immunosorbent assay for testosterone , 1985, Steroids.

[7]  K. Aida,et al.  Endocrine profiles during the short reproductive cycle of an autumn-spawning bitterling, Acheilognathus rhombea. , 1985, General and comparative endocrinology.

[8]  P. Elder,et al.  An enzyme-linked immunosorbent assay (ELISA) for plasma testosterone. , 1985, Journal of steroid biochemistry.

[9]  C. Munro,et al.  Development of a microtitre plate enzyme immunoassay for the determination of progesterone. , 1984, The Journal of endocrinology.

[10]  B. V. Weemen,et al.  Enzyme-immunoassay: a powerful analytical tool. , 1980, Journal of immunoassay.

[11]  H. Hosoda,et al.  The preparation of steroid N-hydroxysuccinimide esters and their reactivities with bovine serum albumin. , 1979, Chemical & pharmaceutical bulletin.

[12]  A. F. Cook,et al.  Daily cycles in serum cortisol levels in the goldfish: effects of photoperiod, temperature, and sexual condition , 1978 .

[13]  P. Samarajeewa,et al.  The radioimmunoassay of steroid glucuronides. The oestrogen C-3 glucuronides as haptens. , 1975, The Biochemical journal.