An ultrasensitive microchip electrophoresis chemiluminescence assay platform for detection of trace biomolecules

An ultrasensitive microchip electrophoresis chemiluminescence (MCE-CL) assay platform based on separation assisted cascade signal amplification was developed for detection of trace biomolecules. In this work, the aptamer was used as a target probe to bind target molecule and triggering cascade signal amplification reaction. The horseradish peroxide labeled DNA (HRP-DNA) was used as signal probe, utilizing nucleic acid hybridization and exonuclease cutting technology realized ultrasensitive detection of biomolecules on the MCE-CL assay platform. Taking gamma interferon (IFN-γ) as a model analyte, the linear range for IFN-γ detection is 8.0 × 10^-15-1.0 × 10^-8 M, the detection limit is 1.6 fM, which is six orders magnitude lower than that of without signal amplification. The proposed method was successfully applied for the quantification of IFN-γ in human plasma samples. It was demonstrated that the MCE-CL assay platform was quick, sensitive, and highly selective. It may serve as a tool for clinical analysis of IFN-γ to assist in the diagnosis of disease.