Highly selective ratiometric electrogenerated chemiluminescence assay of DNA methyltransferase activity via polyaniline and anti-fouling peptide modified electrode

Abstract

In this work, an antifouling electrochemiluminescent (ECL) ratiometric biosensor is designed for the accurate, selective and sensitive detection of DNA methyltransferase (MTase) activity based on a dual-signaling strategy. Briefly, an ITO electrode is used to construct the anti-fouling interfaces with the modification of polyaniline (PANI), AuNPs and peptide. Hairpin DNA molecules containing the symmetric sequence of 5′-CATC-3′ are attached onto the modified ITO electrode and the ds-DNA can be cut off in the presence of Dam MTase and DpnI. The residual DNA and two hairpin DNA could lead to the extension of ds-DNA due to the Hybridization Chain Reaction (HCR). ECL signal is amplified significantly with the insertion of PTC-NH2 molecules into the dsDNA grooves. The ECLPTC-NH2/ECLAu@luminol is found in a logarithmic linear relation with the concentration of Dam MTase. Moreover, owing to the presence of antifouling peptide on the sensing interface, the ECL biosensor was capable of sensing MTase activity in complex biological media, such as FBS samples and human serum with significantly reduced nonspecific adsorption effect. Assaying Dam MTase in complex sample mixture containing 5% calf serum and 5% human serum further proved the feasibility of this ECL biosensor for early clinical diagnosis.