Abstract
Developing accurate and sensitive DNA methyltransferase(MTase)analysis methods is essential for early clinical diagnosis and development of antimicrobial drug targets.In this work,by coupling WO3-x dots-encapsulated metal-organic frameworks(MOFs)as co-reactants and terminal deoxynucleotidyl transferase(TdT)-mediated template-free branched polymerization,a dual signal-amplified electrochemiluminescent(ECL)biosensor was constructed to detect DNA adenine methylation(Dam)MTase.The employment of WO3-x dots-encapsulated MOFs(i.e.,NH2-UIO66@WO3-x)was not only beneficial for biomolecule conju-gation because of the abundant amino groups but also led to a 7-fold enhanced ECL response due to the increased loading of WO3-x.Moreover,TdT-mediated template-free branched polymerization promoted the capture of ECL emitters on the electrode surface,achieving 20-fold enhanced signal amplification.The presented ECL biosensor demonstrated a low detection limit of 2.4 × 10-4 U/mL,and displayed high reli-ability for the detection of Dam MTase in both spiked human serum and E.coli cell samples,and for the screening of potential inhibitors.This study opens a new avenue for designing a dual signal amplification-based ECL bioassay for Dam MTase and screening inhibitors in the fields of clinical diagnosis and drug development.
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