We present herein the very first amperometric biosensor for the quantitative determination of glycine in diverse biological fluids. The biosensor is based on a novel quinoprotein that catalyzes the oxidation of glycine with high specificity. This process is coupled to the redox conversion of Prussian blue in the presence of hydrogen peroxide originating from the enzymatic reaction. The optimized tailoring of the biosensor design consists of the effective encapsulation of the quinoprotein in a chitosan matrix with the posterior addition of an outer Nafion layer, which is here demonstrated to suppress matrix interference. This is particularly important in the case of ascorbic acid, which is known to influence the redox behavior of the Prussian blue. The analytical performance of the biosensor demonstrates fast response time (<7 s), acceptable reversibility, reproducibility, and stability (<6% variation) as well as a wide linear range of response (25-500 μM) that covers healthy (and even most unhealthy) physiological levels of glycine in blood/serum, urine and sweat. A total of 6 real samples from healthy patients and animals were analyzed: two serum, two urine and two sweat samples. The results were validated via commercially available fluorescence kit, displaying discrepancy of less than 9% in all the samples. The unique analytical features and effortless preparation of the new glycine biosensor position it at the forefront of current technologies towards decentralized clinical applications and sport performance monitoring.
Keywords: Biological fluids; Glycine biosensor; Point-of-care-sensing; Prussian blue; Quinoprotein.
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