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A new ‘total’ activin B enzyme‐linked immunosorbent assay (ELISA): development and validation for human samples
Author(s) -
Ludlow Helen,
Phillips David J.,
Myers Michelle,
McLachlan Robert I.,
De Kretser David M.,
Allan Carolyn A.,
Anderson Richard A.,
Groome Nigel P.,
Hyvönen Marko,
Colin Duncan W.,
Muttukrishna Shanthi
Publication year - 2009
Publication title -
clinical endocrinology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.055
H-Index - 147
eISSN - 1365-2265
pISSN - 0300-0664
DOI - 10.1111/j.1365-2265.2009.03567.x
Subject(s) - follicular fluid , medicine , endocrinology , follicular phase , ovarian follicle , andrology , chemistry , biology , embryo , oocyte , microbiology and biotechnology
Summary Background and objective There are currently no sensitive and specific assays for activin B that could be utilized to study human biological fluids. The aim of this project was to develop and validate a ‘total’ activin B ELISA for use with human biological fluids and establish concentrations of activin B in the circulation and fluids from the reproductive organs. Design The new ELISA was validated and then used to measure activin B levels in the circulation of healthy participants, IVF patients, pregnant women and in ovarian follicular fluid and seminal plasma. Patients and measurements Healthy adult subjects ( n = 143), subjects from an IVF clinic ( n = 27) and pregnancy groups ( n = 29) were sampled. Results The sensitivity of the assay was 0·019 ng/ml. Validation of the activin B ELISA showed good recovery (90·7 ± 9·8%) and linearity in biological fluid and cell culture media and low cross‐reactivity with related analytes (inhibin B = 0·077% and activin A = 0·0034%). There was a negative correlation between activin B concentration ( r = −0·281, P < 0·011) and females with increasing age. Patients attending IVF clinics had significantly lower levels of activin B compared with gender‐matched control subjects. Ovarian follicular fluid and seminal plasma had 50–80 fold higher levels of activin B (mean = 5·35 and 3·66 ng/ml respectively) than sera (mean = 0·071 ng/ml). Conclusions This fully validated ELISA for activin B offers a tremendous utility for measuring this protein in a variety of normal physiological processes and in various clinical pathologies.