Development and application of competitive ELISA assays for rat LH and FSH

Rat LH (rLH) and FSH (rFSH) were measured by sensitive and specific competi tion ELISAs. The rat LH ELISA used rLH-I-9 coated plates, an antiserum agai nst rLH and an antibody against rabbit IgG labeled with peroxidase. Using r LH-RP-3 as a standard, rat LH was determined by binding of the anti-LH anti body to rLH-I-9 coated plates. The sensitivity of the assay was 0.8 ng/mL. Similarly, the rat FSH-ELISA used rFSH-I-8 coated plates, an antiserum agai nst rFSH and an antibody against rabbit IgG labeled with peroxidase. Using rFSH-RP-3 as a standard, the FSH-ELISA was also determined by binding of th e anti-FSH antibody to rFSH-I-8 coated plates. The sensitivity of this assa y was 1.25 ng/mL. Both rat LH and FSH ELISA assays are highly specific and provide accurate determination of gonadotrophins in buffers, sera, cell cul ture media, and anterior pituitary extracts. These assays were used for mon itoring the gonadotrophin surge-attenuating factor (GnSAF) and inhibin acti vities present in human follicular fluid (hFF). The 2 new ELISA procedures have practical advantages (safety, convenience, economy) over the RIA metho ds, and they perform as well as the RIA techniques at the same range of con centrations. (C) 1999 by Elsevier Science Inc.