Lactate Dehydrogenase and Glutamate Pyruvate Transaminase biosensing strategies for lactate detection on screen-printed sensors. Catalysis efficiency and interference analysis in complex matrices: from cell cultures to sport medicine

Abstract

Lactate detection with electrochemical biosensors is a fundamental part of the sensing and biosensing research field. Together with glucose detection, this topic attracted scientist attention since the 60s. Lactate is a metabolite involved in many processes among several fields of application, from food degradation, to sport medicine, cell cultures analysis, and clinical monitoring. However, few electrochemical lactate biosensors are currently present in the market.In this work, we analyzed and compared the standard Lactate Dehydrogenase catalysis mechanism with a two-enzymes commercial method that proved to dramatically boost the catalysis efficiency from 0.6% to 72.5%. Both the mechanisms result in the production of reduced Nicotinamide Adenine Dinucleotide (NADH). Therefore, we analyzed its electrochemical detection performance using commercial screen-printed carbon electrodes (SPCE), along with the possible interferences of common mediums and buffers. We found that these sensors can be used for NADH detection down to a 10 μM concentration with a sensitivity of 33.14 nA/μM at a 456 mV overpotential. Moreover, we analytically defined the interferences effects of Dulbecco's Modified Eagle Medium on the sensors by Electrochemical Impedance Spectroscopy measurements and equivalent electrical circuits modeling.Finally, we tested the two-enzyme method in two case studies, i.e., cell cultures lactate monitoring as a valuable tool to evaluate their growth, and lactate quantification in human sweat to monitor physical exercise intensity. We proved the feasibility of the combination of commercial two-enzymes kit and SPCE as a low-cost, accurate, and sensitive lactate biosensor in different applications, regardless of sample interferences.