Cavity Enhanced Immunoassay Measurements in Microtiter Plates using BBCEAS

Zuzana Bajuszova, Zulfiqur Ali, Simon Scott, L. Nitin Seetohul, Meezanul Islam

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Abstract

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.
Original languageEnglish
Pages (from-to)5264-5270
JournalAnalytical Chemistry
Volume88
Issue number10
Early online date27 Apr 2016
DOIs
Publication statusPublished - 17 May 2016

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Absorption spectroscopy
Osteocalcin
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title = "Cavity Enhanced Immunoassay Measurements in Microtiter Plates using BBCEAS",
abstract = "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.",
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Cavity Enhanced Immunoassay Measurements in Microtiter Plates using BBCEAS. / Bajuszova, Zuzana; Ali, Zulfiqur; Scott, Simon; Seetohul, L. Nitin; Islam, Meezanul.

In: Analytical Chemistry, Vol. 88, No. 10, 17.05.2016, p. 5264-5270.

Research output: Contribution to journalArticleResearchpeer-review

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