Blocking-free and self-contained immunoassay platform for one-step point-of-care testing

This paper reports a quantitative and sensitive one-step point-of-care testing (POCT) chip built on a perfluorinated substrate patterned with polydopamine microspots array. The capture antibody was covalently immobilized on the polydopamine microspots, while the fluorescently labelled detection antibody was physically adsorbed on the perfluorinated surface. The POCT chip allowed one-step sandwich immunoassay and was able to directly detect the analytes from the whole blood without sample preprocessing. By further taking advantages of the strong fluorescence quenching ability of the polydopamine, the blocking-free substrate was able to achieve similar performance in detecting and quantifying the protein biomarkers as the substrate with the blocking treatment. The blocking-free strategy not only made the fabrication of the chip simple and convenient, but also improved the chip's sensitivity for biomarker quantification. Finally, we demonstrated that the self-contained POCT platform maintained the performance for one-step immunoassay even after long-term storage. With the POCT platform, we are one step closer to a sample-in-answer-out diagnostic system.