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Cavity-Enhanced Immunoassay Measurements in Microtiter Plates Using BBCEAS
Zuzana Bajuszova1, Zulfiqur Ali1, Simon Scott1
1School of Science and Engineering, Teesside University , Borough Road, Middlesbrough, TS1 3BA, United Kingdom.
Analytical Chemistry
|April 19, 2016
Summary
This study introduces broadband cavity enhanced absorption spectroscopy (BBCEAS) for immunoassays, significantly boosting sensitivity for detecting molecules like osteocalcin. This novel detection method offers a more sensitive and cost-effective approach for various biochemical tests.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Spectroscopy
Background:
- Immunoassays are crucial diagnostic tools, but sensitivity limitations can hinder early disease detection.
- Conventional microplate readers often lack the sensitivity required for detecting low-concentration biomarkers.
- Enhancing detection sensitivity in immunoassays can lead to earlier diagnosis and more cost-effective testing.
Purpose of the Study:
- To report the first detailed application of broadband cavity enhanced absorption spectroscopy (BBCEAS) as a detection system for immunoassays.
- To develop and evaluate a custom-built cavity enhanced microplate reader for enhanced sensitivity measurements.
- To assess the performance of BBCEAS in detecting osteocalcin and a standard colorimetric reaction.
Main Methods:
- Integration of a vertical R ≥ 0.99 optical cavity with a motorized XY stage for 96-well microtiter plates.
- Utilizing a custom-built cavity enhanced microplate reader for measurements.
- Testing with a commercially available osteocalcin sandwich ELISA kit and a standard STREP-HRP colorimetric reaction.
Main Results:
- Achieved a 30-fold increase in path length and a 39-fold increase in measurement sensitivity for osteocalcin detection compared to conventional readers.
- Demonstrated up to a 115-fold increase in sensitivity for a STREP-HRP colorimetric reaction.
- The developed setup showed sensitivity comparable to liquid-phase cavity enhanced studies and approaching fluorometric ELISAs.
Conclusions:
- BBCEAS offers a significant improvement in sensitivity for immunoassay detection.
- The developed cavity enhanced microplate reader provides a powerful tool for biochemical tests relying on absorption detection.
- This technology can enable label-free detection of low-concentration molecules and reduce the cost of diagnostic tests.
