A versatile 3D printed multi-electrode cell for determination of three COVID-19 biomarkers

3D-printing has shown an outstanding performance for the production of versatile electrochemical devices. However, there is a lack of studies in the field of 3D-printed miniaturized settings for multiplex biosensing. In this work, we propose a fully 3D-printed micro-volume cell containing six working electrodes (WEs) that operates with 250 μL of sample. A polylactic acid/carbon black conductive filament (PLA/CB) was used to print the WEs and subsequently modified with graphene oxide (GO), to support protein binding. Cyclic voltammetry was employed to investigate the electrochemical behaviour of the novel multi-electrode cell. In the presence of K₃[Fe(CN)₆], PLA/CB/GO showed adequate peak resolution for subsequent label-free immunosensing. The innovative 3D-printed cell was applied for multiplex voltammetric detection of three COVID-19 biomarkers as a proof-of-concept. The multiple sensors showed a wide linear range with detection limits of 5, 1 and 1 pg mL^-1 for N-protein, S_RBD-protein, and anti-S_RBD, respectively. The sensor performance enabled the selective sequential detection of N protein, S_RBD protein, and anti-S_RBD at biological levels in saliva and serum. In summary, the miniaturized six-electrode cell presents an alternative for the low-cost and fast production of customizable devices for multi-target sensing with promising application in the development of point-of-care sensors.