Development of a two-site solid-phase immunochemiluminescent assay for measurement of dimeric inhibin-A in human serum and other biological fluids.
Inhibin is a heterodimeric glycoprotein that inhibits the secretion of follitropin from the pituitary and has been isolated in two distinct forms composed of a common alpha subunit and either a beta A or beta B subunit. Utilizing paired monoclonal antibodies specific to the alpha and beta A subunit, we have developed an immunochemiluminescent assay for dimeric inhibin-A. The assay is capable of quantifying free and bound inhibin-A in human serum and follicular fluid. The limit of detection is 10 ng/L. Related proteins exhibit little cross-reactivity or interference. Recovery is excellent. Whereas samples from men and postmenopausal women are near the detection limit of the assay, inhibin-A is higher in the luteal than the follicular phase of normally cycling women, 20-fold higher during in vitro fertilization treatment, and approximately 200-fold greater in pregnancy. The assay measures inhibin-A in follicular fluid from a variety of other species.
McConnell
D S
DS
Reproductive Sciences Program, University of Michigan, Ann Arbor 48109-0404, USA. DanMcC@umich.edu
Padmanabhan
V
V
Pollak
T B
TB
Groome
N P
NP
Ireland
J J
JJ
Midgley
A R
AR
D000328:Adult; D000818:Animals; D002417:Cattle; D005260:Female; D005498:Follicular Phase / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D007265:Inhibins / Q000097:blood*; D008163:Luminescent Measurements; D008183:Luteal Phase / Q000097:blood; D046911:Macromolecular Substances; D008297:Male; D017698:Postmenopause / Q000097:blood; D011247:Pregnancy; D011786:Quality Control; D012680:Sensitivity and Specificity; D013045:Species Specificity
D046911:Macromolecular Substances; D007265:Inhibins