Multiplexed analysis of inflammation biomarkers using spectrally-encoded on-chip electrophoresis

We report on development of spectrally multiplexed immunoassays for inflammatory biomarkers utilizing onchip electrophoresis. Direct immunoassays are developed for TNF-alpha and CRP using affinity reagents (antibodies) labeled with spectrally distinct fluorophores. Following off chip incubation, unbound affinity reagents were rapidly resolved from bound immune complex via on-chip electrophoretic separation. Bound immune complex were quantified using 2-color imaging. Dose response curves are generated for both biomarkers at clinically relevant concentrations. With spectral multiplexing, multiple biomarkers can be analyzed in a single microfluidic channel. The technique reported here shows promise as biomarker profiling tool for differential diagnosis in clinical setting.