Electrochemiluminescence and electrochemical dual-mode detection of BACE1 activity based on the assembly of peptide and luminol co-functionalized silver nanoparticles induced by cucurbit[8]uril

Abstract

A novel electrochemiluminescence (ECL) and electrochemical dual-mode sensor was developed for detecting the activity of beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) and screening its inhibitor. Specifically, the adamantane (ADA)-functionalized peptide (P1), a designed substrate peptide for BACE1, was immobilized on the electrode surface via host-guest interaction between β-cyclodextrin (β-CD) and ADA. The aggregation of the peptide (P2) and luminol co-functionalized silver nanoparticles could be induced by cucurbit [8]uril (CB[8] due to the ability of CB[8] to accommodate two aromatic residues simultaneously. The obtained (CB[8]-P2-AgNPs-luminol)n aggregates with both ECL and electrochemical activity, used as the dual-mode signal probe, could be captured to the N-terminal of P1 through CB[8]. Once the substrate P1 was cleaved by BACE1, the probe-binding polypeptide fragment detached from the electrode surface, resulting in a remarkable decrease in the ECL and electrochemical signals. Taking advantage of the signal amplification function of the signal probe, the sensitive dual-mode assay for BACE1 activity can be achieved with the low detection limits of 33.11 pM for ECL and 53.19 pM for electrochemical mode. The superior analytical performance of this novel dual-mode sensor toward BACE1 activity suggested the promising application in early diagnosis of Alzheimer's disease (AD).