Improvements to a surface plasmon resonance-based immunoassay for the steroid hormone progesterone

The effects of modifications to an existing protocol for a surface plasmon resonance biosensor-based inhibition immunoassay for progesterone in cow's milk with a sensitivity of 3.5 ng/mL were examined to establish if the detection limit could be further reduced to broaden the potential applicat...

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Bibliographic Details
Published in:Journal of AOAC International 2006-05, Vol.89 (3), p.838-842
Main Authors: Gillis, E.H, Traynor, I, Gosling, J.P, Kane, M
Format: Article
Language:English
Subjects:
animal fertility
Animals
Biosensing Techniques - methods
biosensors
Cattle
dairy cows
Dose-Response Relationship, Drug
estrous cycle
food analysis
Hormones
Hormones, Sex
Immunoassay - methods
immunoassays
Milk
Progesterone
Progesterone - analysis
quantitative analysis
rapid methods
Steroid hormones
Steroids - analysis
surface plasmon resonance
Surface Plasmon Resonance - instrumentation
Surface Plasmon Resonance - methods
Temperature
Time Factors
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Summary:The effects of modifications to an existing protocol for a surface plasmon resonance biosensor-based inhibition immunoassay for progesterone in cow's milk with a sensitivity of 3.5 ng/mL were examined to establish if the detection limit could be further reduced to broaden the potential applications of the assay. The mean relative standard deviation of duplicate measurements was 0.62% and the high precision resulted in very low values for the lower detection limits. Hence, the standard concentrations giving 95% maximum binding [effective dose (ED 95)] were compared instead. The ED 95 was not affected within a running temperature range of 20-37 degrees C, or at a flow rate and a contact time above 20 microliter/min and 90 s, respectively. Increasing both the absolute sample volume and the antibody dilution improved sensitivity. However, there was a simultaneous reduction in the working range when the assay was applied to milk due to nonspecific binding. Less antibody was associated with large decreases in the maximum binding, but because the high precision extended over a broad analytical range, an ED 95 of 0.4-0.6 ng/mL in milk and 35-60 pg/mL in HBS-EP buffer were achieved. Thus, simple procedural modifications with the same sensor chip can alter performance characteristics of the assay as required for different applications.
ISSN:1060-3271
1944-7922
DOI:10.1093/jaoac/89.3.838