3D printed enzymatic microchip for multiplexed electrochemical biosensing

Abstract The low-cost e-fabrication of specialized multianalyte biosensors within the point-of-care (POC) settings in a few minutes remains a great challenge. Unlike prefabricated biosensors, 3D printing seems to be able to meet this challenge, empowering the end user with the freedom to create on-demand devices adapted to immediate bioanalytical need. Here, we describe a novel miniature all-3D-printed 4-electrode biochip, capable of the simultaneous determination of different biomarkers in a single assay. The chip is utterly fabricated via an one-step 3D printing process and it is connected to a mini portable bi-potentiostant, permitting simultaneous measurements. The bioanalytical capability of the microchip is demonstrated through the simultaneous amperometric determination of two cardiac biomarkers (cholesterol and choline) in the same blood droplet, via enzymatic assays developed on its two tiny integrated electrodes. The simultaneous determination of cholesterol and choline is free from cross-talk phenomena and interferences offering limits of detection much lower than the cut-off levels of these biomarkers in blood for coronary syndromes. The biodevice is an easy-constructed, low-cost, sensitive and e-transferable POC chip with wide scope of applicability to other enzymatic bioassays.