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Real-Time Highly-Sensitive Protein Quantification Through On-Chip Chemiluminescence

preprint
submitted on 13.07.2020 and posted on 14.07.2020 by Hoi Kei Chiu, Tadas Kartanas, Sean R. A. Devenish, Kadi Saar, Carina Mouritsen Luxhøj, Tuomas Knowles

A range of experimental methods have been developed to achieve highly sensitive detection and quantification of proteins. The majority of these methods rely on fluorescence-mediated readouts and, as such, their sensitivity can be affected by factors such as photobleaching of fluorophores and background signal from the illumination source. Both of these limitations can be overcome by using chemiluminescence-based detection: in contrast to fluorescence, chemiluminescence can be generated in an excitation source free manner, which allows for a significant reduction in background noise and for the use of an optical setup that comprises only a detection element. Here, we develop a highly-sensitive protein quantification platform by combining chemiluminescent detection of proteins with microfluidic mixing and detection. We use the platform to demonstrate quantitative detection of proteins over a concentration range of five orders of magnitude

and down to 10 pg mL−1, corresponding to pM concentrations. Owing to the general presence of amine groups in peptides and proteins, our demonstrated system is applicable to characterising any protein sample and it can be used to quantify unlabelled samples.

Funding

Engineering and Physical Sciences Research Council

Schmidt Science Fellowship programme in partnership with the Rhodes Trust

European Research Council under the European Union’s Seventh Framework Programme

Newman Foundation

History

Email Address of Submitting Author

kls78@cam.ac.uk

Institution

University of Cambridge

Country

United Kingdom

ORCID For Submitting Author

0000-0002-5926-3628

Declaration of Conflict of Interest

The authors declare no conflict of interest

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