Multiplexed and simultaneous biosensing in a 3D-printed portable six-well smartphone operated electrochemiluminescence standalone point-of-care platform.

3D-printed portable devices have immense and proven potential to transform the field of electrochemiluminescence (ECL) for diverse biochemical applications. 3D printing (3DP) offers unparalleled ability to build tiny devices in a single step with high accuracy and compatibility, and integrability as per the requirement. In this study, for the first time, a six-well 3D-printed closed bipolar electrochemiluminescence (3DP-CBPE-ECL) device has been successfully fabricated and validated by performing single-step detection of various biochemicals such as glucose and choline. Luminol/H2O2-based enzymatic reactions were performed with optimized parameters for selective sensing of glucose and choline. The single-step detection of glucose and choline was accomplished for the linear ranges of 0.1 to 10 mM and 0.1 to 5 mM, with a limit of detections (LODs) of 24 µM and 10 µM, respectively. A smartphone was leveraged to execute multiple activities such as powering the ECL device, capturing ECL images, and calculating the ECL intensity of the obtained ECL signal. The feasibility of a six-well 3DP-CBPE-ECL device was tested by sensing glucose and choline simultaneously in a single device at three different concentrations. Furthermore, the concentration of glucose and choline was calculated in real blood serum using the conventional additive (spiking) method, demonstrating the high practicability of the fabricated ECL device and yielding promising findings. Finally, based on the obtained results and other advantages such as low-cost, fast prototyping and requirement of a minimum sample volume, the fabricated six-well 3DP-CBPE-ECL device has shown potential to be used in the field of biochemical applications.