Cavity enhanced immunoassay measurements in microtiter plates using BBCEAS

Bajuszova, Z., Ali, Z., Scott, S.M., Seetohul, L.N. ORCID: 0000-0001-6261-0259 and Islam, M., 2016. Cavity enhanced immunoassay measurements in microtiter plates using BBCEAS. Analytical Chemistry, 88 (10), pp. 5264-5270. ISSN 0003-2700

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We report on the first detailed use of broadband cavity enhanced absorption spectroscopy (BBCEAS) as a detection system for immunoassay. A vertical R ≥ 0.99 optical cavity was integrated with a motorised XY stage, which functioned as a receptacle for 96 well microtiter plates. The custom built cavity enhanced microplate reader was used to make measurements on a commercially available osteocalcin sandwich ELISA kit. A 30 fold increase in path length was obtained with a minimum detectable change in the absorption coefficient, αmin(t), of 5.3 × 10-5 cm-1 Hz-1/2. This corresponded to a 39 fold increase in the sensitivity of measurement when directly compared to measurements in a conventional microplate reader. Separate measurements of a standard STREP-HRP colorimetric reaction in microtiter plates of differing optical quality produced an increase in sensitivity of up to 115 fold compared to a conventional microplate reader. The sensitivity of the developed setup compared favorably with previous liquid-phase cavity enhanced studies and approaches the sensitivity of typical fluorometric ELISAs. It could benefit any biochemical test which uses single pass absorption as a detection method, through either the label free detection of biologically important molecules at lower concentrations or the reduction in the amount of expensive biochemicals needed for a particular test, leading to cheaper tests.

Item Type: Journal article
Publication Title: Analytical Chemistry
Creators: Bajuszova, Z., Ali, Z., Scott, S.M., Seetohul, L.N. and Islam, M.
Publisher: ACS (American Chemical Society) Publications
Date: 18 April 2016
Volume: 88
Number: 10
ISSN: 0003-2700
Divisions: Schools > School of Science and Technology
Record created by: Jill Tomkinson
Date Added: 26 Apr 2016 13:17
Last Modified: 13 Oct 2017 10:52

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