Label Free Detection of L-Glutamate Using Microfluidic Based Thermal Biosensor

Bioengineering (Basel). 2015 Jan 12;2(1):2-14. doi: 10.3390/bioengineering2010002.

Abstract

A thermoelectric biosensor for the detection of L-glutamate concentration was developed. The thermoelectric sensor is integrated into a micro-calorimeter which measures the heat produced by biochemical reactions. The device contains a single flow channel that is 120 µm high and 10 mm wide with two fluid inlets and one fluid outlet. An antimony-bismuth (Sb-Bi) thermopile with high common mode rejection ratio is attached to the lower channel wall and measures the dynamic changes in the temperature when L-glutamate undergoes oxidative deamination in the presence of glutamate oxidase (GLOD). The thermopile has a Seebeck coefficient of ~7 µV·(m·K)-1. The device geometry, together with hydrodynamic focusing, eliminates the need of extensive temperature control. Layer-by-layer assembly is used to immobilize GLOD on the surface of glass coverslips by alternate electrostatic adsorption of polyelectrolyte and GLOD. The impulse injection mode using a 6-port injection valve minimizes sample volume to 5 µL. The sensitivity of the sensor for glutamate is 17.9 nVs·mM-1 in the linear range of 0-54 mM with an R² value of 0.9873. The lowest detection limit of the sensor for glutamate is 5.3 mM.

Keywords: L-glutamate; L-glutamate oxidase; biosensor; label-free; layer-by-layer self-assembly; thermoelectric.