Autonomous electrochemical biosensing of glial fibrillary acidic protein for point-of-care detection of central nervous system injuries.

The integration of electrochemical biosensors into fluid handling units such as paper-based, centrifugal, and capillary microfluidic devices has been explored with the purpose of developing point-of-care platforms for quantitative detection of bodily fluid markers. However, the present fluidic device designs largely lack the capacity of full assay automation, needing manual loading of one or multiple reagents or requiring external devices for liquid manipulation. Such fluidic handing platforms also require universality for detecting various biomarkers. These platforms are also largely produced using materials unsuitable for scalable manufacturing and with a high production cost. The mechanism of fluid flow also often induces noise to the embedded biosensors which adversely impacts the accuracy of biosensing. This work addresses these challenges by presenting a reliable design of a fully automated and universal capillary-driven microfluidic platform that automates several steps of label-free electrochemical biosensing assays. These steps include sample aliquoting, controlled incubation, removal of non-specific bindings, reagent mixing and delivery to sensing electrodes, and electrochemical detection. The multilayer architecture of the microfluidic device is made of polymeric and adhesive materials commercially used for the fabrication of point-of-care devices. The design and geometry of different components of the device (, sampling unit, mixer, resistances, delay valves, interconnecting components) were optimized using a combined experimental testing and numerical fluid flow modeling to reach high reproducibility and minimize the noise-induced to the biosensor. As a proof of concept, the performance of this on-chip immunosensing platform was demonstrated for rapid and autonomous detection of glial fibrillary acidic proteins (GFAP) in phosphate-buffered saline (PBS). The microfluidic immunosensing device exhibited a linear detection range of 10-1000 pg mL for the detection of GFAP within 30 min, with a limit of detection (LoD) and sensitivity of 3 pg mL and 39 mL pg mm in PBS, respectively. Owing to its simplicity, sample-to-result performance, universality for handing different biofluids, low cost, high reproducibility, compatibility with scalable production, and short analysis time, the proposed biosensing platform can be further adapted for the detection of other biomarkers in different clinical bodily fluids for rapid diagnostic and prognostic applications.