Real-time, continuous detection of maltose using bioluminescence resonance energy transfer (BRET) on a microfluidic system

Le, N, Gel, M, Zhu, Y, Dacres, H, Anderson, A and Trowell, S 2014, 'Real-time, continuous detection of maltose using bioluminescence resonance energy transfer (BRET) on a microfluidic system', Biosensors and Bioelectronics, vol. 62, pp. 177-181.


Document type: Journal Article
Collection: Journal Articles

Title Real-time, continuous detection of maltose using bioluminescence resonance energy transfer (BRET) on a microfluidic system
Author(s) Le, N
Gel, M
Zhu, Y
Dacres, H
Anderson, A
Trowell, S
Year 2014
Journal name Biosensors and Bioelectronics
Volume number 62
Start page 177
End page 181
Total pages 5
Publisher Elsevier
Abstract We have previously shown that a genetically encoded bioluminescent resonance energy transfer (BRET) biosensor, comprising maltose binding protein (MBP) flanked by a green fluorescent protein (GFP 2 ) at the N-terminus and a variant of Renilla luciferase (RLuc2) at the C-terminus, has superior sensitivity and limits of detection for maltose, compared with an equivalent fluorescent resonance energy transfer (FRET) biosensor. Here, we demonstrate that the same MBP biosensor can be combined with a microfluidic system for detection of maltose in water or beer. Using the BRET-based biosensor, maltose in water was detected on a microfluidic chip, either following a pre-incubation step or in real-time with similar sensitivity and dynamic range to those obtained using a commercial 96-well plate luminometer. The half-maximal effective concentrations (EC 50 ) were 2.4×10 -7 M and 1.3×10 -7 M for maltose detected in pre-incubated and real-time reactions, respectively. To demonstrate real-time detection of maltose in a complex medium, we used it to estimate maltose concentration in a commercial beer sample in a real-time, continuous flow format. Our system demonstrates a promising approach to in-line monitoring for applications such as food and beverage processing.
Subject Manufacturing Engineering not elsewhere classified
Keyword(s) Beer
Bioluminescent resonance energy transfer (BRET)
In-line monitoring
Maltose
Microfluidics
Real-time
DOI - identifier 10.1016/j.bios.2014.06.032
Copyright notice Crown Copyright © 2014 Published by Elsevier B.V. All rights reserved.
ISSN 0956-5663
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