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

• Published in: Biosensors & bioelectronics Vol. 62; pp. 177 - 181
• Main Authors: Le, Nam Cao Hoai, Gel, Murat, Zhu, Yonggang, Dacres, Helen, Anderson, Alisha, Trowell, Stephen C.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 15.12.2014; Elsevier
• Subjects: Beer; Beer - analysis; Biological and medical sciences; Bioluminescence Resonance Energy Transfer Techniques - methods; Bioluminescent resonance energy transfer (BRET); Biosensing Techniques - methods; Biosensors; Biotechnology; Computer Systems; Dynamical systems; Energy transfer; Fundamental and applied biological sciences. Psychology; Green Fluorescent Proteins; In-line monitoring; Luciferases, Renilla; Maltose; Maltose - analysis; Maltose-Binding Proteins; Microfluidic Analytical Techniques - methods; Microfluidics; Proteins; Real time; Renilla; Water; Bioluminescent resonance energy transfer (BRET); Maltose; In-line monitoring; Real-time; Microfluidics; Beer; Fluid mechanics; Bioluminescence; Resonance energy transfer; Real time; Continuous process; Resonant energy transfer; Miniaturization; Detection; Monitoring
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2014.06.032

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 (GFP2) 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 (EC50) were 2.4×10−7M and 1.3×10−7M 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. •A BRET2-based biosensor was used in real-time and continuous microfluidic detection.•Maltose was accurately measured in water and in beer using the biosensor.•EC50 was 2.4×10−7M for maltose detected in pre-incubated assay.•For the real-time assay, EC50 was 1.3×10−7M.•Potential application in in-line monitoring in the food and beverage industries.