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..
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 language | English |
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Pages (from-to) | 867-73 |
Number of pages | 7 |
Journal | Clinical Endocrinology |
Volume | 71 |
Issue number | 6 |
DOIs | |
Publication status | Published - Dec 2009 |