Designing electrochemical microfluidic multiplexed biosensors for on-site applications

Clinical assessment based on a single biomarker is in many circumstances not sufficient for adequate diagnosis of a disease or for monitoring its therapy. Multiplexing, the measurement of multiple analytes from one sample and/or of the same target from different samples simultaneously, could enhance the accuracy of the diagnosis of diseases and their therapy success. Thus, there is a great and urgent demand for multiplexed biosensors allowing a low-cost, easy-to-use, and rapid on-site testing. In this work, we present a simple, flexible, and highly scalable strategy for implementing microfluidic multiplexed electrochemical biosensors (BiosensorX). Our technology is able to detect 4, 6, or 8 (different) analytes or samples simultaneously using a sequential design concept: multiple immobilization areas, where the assay components are adsorbed, followed by their individual electrochemical cells, where the amperometric signal readout takes place, within a single microfluidic channel. Here, first we compare vertical and horizontal designs of BiosensorX chips using a model assay. Owing to its easier handling and superior fluidic behavior, the vertical format is chosen as the final multiplexed chip design. Consequently, the feasibility of the BiosensorX for multiplexed on-site testing is successfully demonstrated by measuring meropenem antibiotics via an antibody-free beta-lactam assay. The multiplexed biosensor platform introduced can be further extended for the simultaneous detection of other anti-infective agents and/or biomarkers (such as renal or inflammation biomarkers) as well as different (invasive and non-invasive) sample types, which would be a major step towards sepsis management and beyond.

Automated summary

Clinical assessment often requires the measurement of multiple analytes or samples to improve the accuracy of diagnosis and therapy. In this study, a microfluidic multiplexed electrochemical biosensor technology (BiosensorX) was developed, allowing simultaneous detection of multiple analytes or samples within a single microfluidic channel. The vertical design of the BiosensorX chip was chosen for its ease of use and superior fluidic behavior. The feasibility of the BiosensorX for on-site testing was demonstrated with the measurement of meropenem antibiotics using an antibody-free beta-lactam assay. This multiplexed biosensor platform can be further extended for the detection of other anti-infective agents and biomarkers, as well as different sample types.