Electrochemiluminescence (ECL) biosensor based on tris(2,2'-bipyridyl)ruthenium(II) with glucose and lactate dehydrogenases encapsulated within alginate hydrogels

Abstract

Two dehydrogenase enzymes (glucose, GDH, and lactate, LDH, dehydrogenases) encapsulated within alginate hydrogels were deposited on glassy carbon electrodes. The as-prepared enzyme modified alginate hydrogels were utilized as electrochemiluminescence (ECL)-based biosensors for the indirect detection of glucose and lactic acid upon reaction between NADH and tris(2,2'-bipyridyl) ruthenium (II) [Ru(bpy)₃]²⁺. The ECL response was obtained from the redox reaction between the substrate, the cofactor NAD⁺ and the encapsulated enzyme. The production of NADH resulting from the enzymatic reaction led to the ECL emission upon reaction with [Ru(bpy)₃]²⁺. The biosensors showed good stability and repeatability, with linear range between 0.56 and 4.2 µM and limit of detection of 0.84 µM for glucose, and linear range between 5 and 30 µM with a limit of detection of 2.52 µM for lactic acid. These ECL-based biosensors showed good sensitivity when tested in the presence of common interfering species. These biosensors were utilized in artificial sweat and were characterized by good reproducibility and repeatability. The results herein presented suggest that the dehydrogenases encapsulated within alginate hydrogels have potential for the development of biocompatible sensors for detection of glucose and lactic acid in physiological fluids.

Keywords: Bioelectrochemistry; Biosensors; Electrochemiluminescence; Enzymes; Glucose dehydrogenase; Lactate dehydrogenase.