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

Autor: Ludlow H; Centre for Proteins and Peptides, School of Life Sciences, Oxford Brookes University, Headington, Oxford, UK. helenludlow9@googlemail.com, Phillips DJ, Myers M, McLachlan RI, de Kretser DM, Allan CA, Anderson RA, Groome NP, Hyvönen M, Duncan WC, Muttukrishna S
Jazyk: angličtina
Zdroj: Clinical endocrinology [Clin Endocrinol (Oxf)] 2009 Dec; Vol. 71 (6), pp. 867-73. Date of Electronic Publication: 2009 May 28.
DOI: 10.1111/j.1365-2265.2009.03567.x
Abstrakt: 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.
Databáze: MEDLINE
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