Sub-nanomolar detection of thrombin activity on a microfluidic chip

Le, N, Gel, M, Zhu, Y, Wang, J, Dacres, H, Anderson, A and Trowell, S 2014, 'Sub-nanomolar detection of thrombin activity on a microfluidic chip', Biomicrofluidics, vol. 8, no. 6, pp. 1-12.


Document type: Journal Article
Collection: Journal Articles

Title Sub-nanomolar detection of thrombin activity on a microfluidic chip
Author(s) Le, N
Gel, M
Zhu, Y
Wang, J
Dacres, H
Anderson, A
Trowell, S
Year 2014
Journal name Biomicrofluidics
Volume number 8
Issue number 6
Start page 1
End page 12
Total pages 12
Publisher A I P Publishing LLC
Abstract Bioluminescence resonance energy transfer (BRET) is a form of Förster resonance energy transfer. BRET has been shown to support lower limits of detection than fluorescence resonance energy transfer (FRET) but, unlike FRET, has not been widely implemented on microfluidic devices for bioanalytical sensing. We recently reported a microscope-based microfluidic system for BRET-based biosensing, using a hybrid, high quantum-efficiency, form of BRET chemistry. This paper reports the first optical fiber-based system for BRET detection on a microfluidic chip, capable of quantifying photon emissions from the low quantum-efficiency BRET2 system. We investigated the effects of varying core diameter and numerical aperture of optical fibers, as well as varying microfluidic channel design and measurement conditions. We optimized the set-up in order to maximize photon counts and minimize the response time. The optimized conditions supported measurement of thrombin activity, with a limit of detection of 20 pM, which is lower than the microscope-based system and more than 20 times lower than concentrations reported to occur in plasma clots.
Subject Medical Biotechnology not elsewhere classified
Keyword(s) Resonance Energy-Transfer
Protein-Protein Interactions
Transfer Bret
Fluorescent Protein
Renilla Luciferase
Biosensors
System
Assay
FRET
DOI - identifier 10.1063/1.4902908
Copyright notice © 2014 AIP Publishing LLC.
ISSN 1932-1058
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