Magnetic Control of an Electrochemical Microfluidic Device with an Arrayed Immunosensor for Simultaneous Multiple Immunoassays

Abstract

Abstract Background: Methods based on magnetic bead probes have been developed for immunoassay, but most involve complicated labeling or stripping procedures and are unsuitable for routine use. Methods: We synthesized magnet core/shell NiFe2O4/SiO2 nanoparticles and fabricated an electrochemical magnetic controlled microfluidic device for the detection of 4 tumor markers. The immunoassay system consisted of 5 working electrodes and an Ag/AgCl reference electrode integrated on a glass substrate. Each working electrode contained a different antibody immobilized on the NiFe2O4/SiO2 nanoparticle surface and was capable of measuring a specific tumor marker using noncompetitive electrochemical immunoassay. Results: Under optimal conditions, the multiplex immunoassay enabled the simultaneous detection of 4 tumor markers. The sensor detection limit was <0.5 μg/L (or <0.5 kunits/L) for most analytes. Intra- and interassay imprecisions (CVs) were <4.5% and 8.7% for analyte concentrations >5 μg/L (or >5 kunits/L), respectively. No nonspecific adsorption was observed during a series of procedures to detect target proteins, and electrochemical cross-talk (CV) between neighboring sites was <10%. Conclusion: This immunoassay system offers promise for label-free, rapid, simple, cost-effective analysis of biological samples. Importantly, the chip-based immunosensor could be suitable for use in the mass production of miniaturized lab-on-a-chip devices and open new opportunities for protein diagnostics and biosecurity.