Design of a photoelectrochemical lab-on-a-chip immunosensor based on enzymatic production of quantum dots in situ

In this work we report the development and validation of a photoelectrochemical immunosensor on the basis of alkaline phosphatase (ALP)-linked immunoassay for the detection of human serum albumin as a model analyte. In this biosensor, oriented immobilization of capture antibodies on aminated polystyrene was achieved via physical adsorption. After the interaction with the analyte, ALP immobilised on the surface through the sandwich immunoassay catalyses the hydrolysis of sodium thiophosphate (TP) to hydrogen sulphide (H₂S) which in the presence of cadmium ions yields CdS quantum dots (QDs). The electrical current is generated in the course of the photoelectrochemical process (PEC) during irradiation of the CdS QDs with a UV LED (365 nm) on home-made screen-printed carbon electrodes modified with a conductive polymer. Reaction time, steps and volumes were optimized for the miniaturization of the process in order to develop a lab-on-a-chip platform. The microfluidic system was designed with optimised parameters to fabricate the immunosensor combining the immunoassay with PEC detection. The final system presents a sensitivity comparable to that of the commercial kit thanks to the signal amplification enabled by the enzymatic growth of CdS QDs in situ. This photoelectrochemical immunosensing strategy potentially opens up a new avenue for the detection of a wide range of analytes of interest due to the universal and effective enzymatic signal amplification method. Moreover, the developed bioanalytical device allows for a great reduction of time and reagents compared to exiting commercial assays, making it suitable for point-of-care applications.