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 mu L 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-ofconcept. The multiple sensors showed a wide linear range with detection limits of 5, 1 and 1 pg mL- 1 for Nprotein, SRBD-protein, and anti-SRBD, respectively. The sensor performance enabled the selective sequential detection of N protein, SRBD protein, and anti-SRBD 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.

Automated summary

This study presents a fully 3D-printed micro-volume cell with six working electrodes, suitable for multiplex biosensing. The use of a PLA/CB conductive filament modified with GO allows for label-free immunosensing. The 3D-printed cell was successfully applied for the selective detection of COVID-19 biomarkers in saliva and serum, making it a promising tool for point-of-care sensors.