Quantitative Detection of Digoxin in Plasma Using Small-Molecule Immunoassay in a Recyclable Gravity-Driven Microfluidic Chip.

Immunoassays are critical for clinical diagnostics and biomedical research. However, two major challenges remaining in conventional immunoassays are precise quantification and development of immunoassays for small-molecule detection. Here, a two signal-mode small-molecule immunoassay containing an internal reference that provides high stability and reproducibility compared to conventional small-molecule immunoassays is presented. A system is developed for quantitative monitoring of the digoxin concentration in plasma in the clinically relevant range (0.6-2.6 nm). Furthermore, the model system is integrated into a simple gravity-driven microfluidic chip (G-Chip) requiring only 10 µL plasma. The G-Chip allows fast detection without any complex operation and can be recycled for at least 50 times. The assay, and the G-Chip in particular, has the potential for further development of point-of-care (POC) diagnostics.