Battery-triggered microfluidic paper-based multiplex electrochemiluminescence immunodevice based on potential-resolution strategy.

A potential-resolution strategy for multiplex electrochemiluminescence (ECL) immunoassay on a microfluidic paper-based analytical device (μ-PAD) was demonstrated for the first time, using tris-(bipyridine)-ruthenium(ii) (Ru(bpy)(3)(2+)) and carbon nanodots (CNDs) as the ECL labels for high-throughput ECL immunoassay on μ-PAD. Based on this strategy, simultaneous detection of four tumor markers using only two screen-printed carbon working electrodes (one electrode for two analytes) on an immunodevice consisting of a piece of patterned paper (denoted as μ-PECLI in this work) was realized, which is a simplification of the configuration of traditional μ-PADs. As a further development of μ-PECLI in low-cost and disposable applications, battery-triggered (constant-potential) ECL detection on μ-PECLI was developed, allowing the traditional electrochemical workstation to be abandoned. In addition, to exactly control the output voltage of the battery, a low-cost and simple voltage-controller was designed and fabricated for the first time. The battery-triggered ECL immunoassay principle on μ-PECLI is explained. We found that the simultaneous determination of two analytes in one paper working zone could be achieved by controlling the operational constant-potential (+1.2 V for Ru(bpy)(3)(2+) labels and -1.2 V for CNDs labels (vs. Ag/AgCl auxiliary electrode)) by simply reversing the connection mode. Finally, four tumor markers in human serum samples from the Tumor Hospital were assayed and the results obtained were in acceptable agreement with the reference values from parallel single-analyte test. This battery-triggered μ-PECLI provides a new strategy for high-throughput, low-cost, sensitive, automated and simultaneous multiplex immunoassay and point-of-care diagnosis.