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Dysregulations in metabolic and physiological cellular processes are generally accompanied by changes in concentration profiles of multiple biomarkers. Measuring dynamic changes in concentrations of several affected biomarkers can enhance our comprehension of the underlying disease pathophysiology and/or mechanisms of cytotoxicity. Most analytical detection techniques require samples to be obtained at pre-defined times for offline analysis. However, the inadequate temporal resolution of these methods can lead to key biological events that occur at shorter timescales, being overlooked. Electrochemical biosensors are relatively simple, cost-effective detection techniques that have been used extensively for continuous analyte monitoring, and more recently, for sensing multiple analytes. We describe the design and functionality of a novel electrochemical detection platform, comprising a screen-printed Pt electrode array, a microfluidic chip, and an automated pump and valve system, capable of quantitatively detecting up to eight analytes. As a proof of concept, we show the simultaneous, sensitive, and selective amperometric detection of four essential biomarkers- glucose, lactate, glutamate, and acetylcholine, in real time. We also demonstrate the biological application of this platform by continuously monitoring exocytotic glutamate release from human induced pluripotent stem cell-derived neuronal cultures over time. Excessive accumulation of glutamate in the synaptic cleft leads to prolonged stimulation of glutamate receptors on neurons, triggering a cascade of neurotoxic events that ultimately results in loss of neuronal function and cell death. The results suggest that our multianalyte electrochemical sensor platform can be coupled with more complex biological systems such as in vitro organotypic models, for the accurate monitoring of biomarker changes during adverse events like acute neurotoxicity and microbial infection. |
