Immunosensor for Detection of Inhibitory Neurotransmitter gamma-Aminobutyric Acid Using Quartz Crystal Microbalance

摘要

A piezoelectric immunosensor for sensing the low molecular weight neurotransmitter gamma-aminobutyric acid (GABA), one of two major inhibitory neurotransmitters in the central nervous system, is described. The sensing interface consists of a dextran layer covalently attached to a self-assembled monolayer of thiolamine compound on the surface of gold electrodes of the crystals. The dextran layer is further modified with GABA molecules to act as the biosensing layer. The affinity binding of monoclonal anti-GABA antibody on the modified piezoelectric crystals is studied in real time without any additional labels. The equilibrium association constant, K_(eq) for binding between anti-GABA antibody and GABA molecules is 14.5 (mu)g(centre dot)mL~(-1). The detection limit for anti-GABA is approx10 nM. The sensitivity of the sensor at a concentration corresponding to half-maximal response is 13.6 ng/mL(centre dot)Hz. The functionalized sensor substrate is subsequently used for competitive determination of different concentrations of free GABA (range of 5 (mu)M-50 mM) in PBS-BSA buffer. The detection limit of the immunosensor for sensing GABA with maximum sensitivity is approx42 (mu)M.