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


METHODS
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.


METHODS
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.
Ludlow
Helen
H
Centre for Proteins and Peptides, School of Life Sciences, Oxford Brookes University, Headington, Oxford, UK. helenludlow9@googlemail.com
Phillips
David J
DJ
Myers
Michelle
M
McLachlan
Robert I
RI
de Kretser
David M
DM
Allan
Carolyn A
CA
Anderson
Richard A
RA
Groome
Nigel P
NP
Hyvönen
Marko
M
Duncan
W Colin
WC
Muttukrishna
Shanthi
S
D028341:Activins / Q000032:analysis*; D000293:Adolescent; D000328:Adult; D000368:Aged; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015571:Follicular Fluid / Q000737:chemistry; D006801:Humans; D008297:Male; D008875:Middle Aged; D011247:Pregnancy; D012661:Semen / Q000737:chemistry; D055815:Young Adult
C044147:activin B; D028341:Activins