A new 'total' activin B enzyme-linked immunosorbent assay (ELISA): development and validation for human samples

Helen Ludlow, David J Phillips, Michelle Myers, Robert I McLachlan, David M de Kretser, Carolyn A Allan, Richard A Anderson, Nigel P Groome, Marko Hyvönen, W Colin Duncan, Shanthi Muttukrishna

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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 center dot 019 ng/ml. Validation of the activin B ELISA showed good recovery (90 center dot 7 +/- 9 center dot 8%) and linearity in biological fluid and cell culture media and low cross-reactivity with related analytes (inhibin B = 0 center dot 077% and activin A = 0 center dot 0034%). There was a negative correlation between activin B concentration (r = -0 center dot 281, P < 0 center dot 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 center dot 35 and 3 center dot 66 ng/ml respectively) than sera (mean = 0 center dot 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..
Original languageEnglish
Pages (from-to)867-73
Number of pages7
JournalClinical Endocrinology
Volume71
Issue number6
DOIs
Publication statusPublished - Dec 2009

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