Electrochemiluminescence Biosensor Based on 3-D DNA Nanomachine Signal Probe Powered by Protein-Aptamer Binding Complex for Ultrasensitive Mucin 1 Detection

Abstract

Herein, we fabricated a novel electrochemiluminescence (ECL) biosensor for ultrasensitive detection of mucin 1 (MUC1) based on a three-dimensional (3-D) DNA nanomachine signal probe powered by protein-aptamer binding complex. The assembly of 3-D DNA nanomachine signal probe achieved the cyclic reuse of target protein based on the protein-aptamer binding complex induced catalyzed hairpin assembly (CHA), which overcame the shortcoming of protein conversion with enzyme cleavage or polymerization in the traditional examination of protein. In addition, CoFeO, a mimic peroxidase, was used as the nanocarrier of the 3-D DNA nanomachine signal probe to catalyze the decomposition of coreactant HO to generate numerous reactive hydroxyl radical OH as the efficient accelerator of N-(aminobutyl)-N-(ethylisoluminol) (ABEI) ECL reaction to amplify the luminescence signal. Simultaneously, the assembly of 3-D DNA nanomachine signal probe was executed in solution, which led to abundant luminophore ABEI be immobilized around the CoFeO surface with amplified ECL signal output since the CHA reaction was occurred unencumberedly in all directions under homogeneous environment. The prepared ECL biosensor showed a favorable linear response for MUC1 detection with a relatively low detection limit of 0.62 fg mL. With excellent sensitivity, the strategy may provide an efficient method for clinical application, especially in trace protein determination.

Label-free sensing of the binding state of MUC1 peptide and anti-MUC1 aptamer solution in fluidic chip by terahertz spectroscopy

“…Aptamers, single-stranded RNA or DNA oligonucleotide selected by the systematic evolution of ligands by exponential enrichment (SELEX) methodology, interact with targets by forming special three-dimensional conformations. Since the concept of aptamer was first proposed by Ellington in 1990 [7], it has been presented as a regnant bioreceptor for the construction of various aptasensors to detect ions [8,9], proteins [1,10], and cells [11,12], as aptamers possess high affinity, exquisite specificity, reduced immunogenicity, increased stability, and good reproducibility. Moreover, aptamers could be massively synthesized and easily modified via chemical processes, which is more costeffective [13].…”

“…Subsequently, an aptamer (anti-MUC1 S2.2) has been confirmed as a functional molecule, specifically binding to a common sequence (APDTRPAPG) of the VTR region of MUC1. To date, this MUC1 and aptamer binding reaction has led to discerning diagnostic aptasensors for detecting and monitoring tumor progression [1,[10][11][12]. In particular, the presence of divalent metal ions (e.g., Mg 2+ , Ca 2+ ) was indispensable for an aptamer to form the characteristic secondary structure, contributing to the selection process of aptamers and regeneration of aptasensors, in contrast to most existing biosensors [11].…”

Programmable and Multifunctional DNA‐Based Materials for Biomedical Applications

“…In order to direct capture and detect liver cancer exosomes, Wang et al integrated the aptamer technology, DNA‐based nanostructure, and portable electrochemical devices to fabricate a nanotetrahedron‐assisted aptasensor, which increased 100‐fold sensitivity compared to the single‐stranded aptamer‐functionalized aptasensor ( Figure a) . The ultrasensitive mucin 1 detection with a detection limit of 0.62 fg mL −1 was achieved by using the electro‐chemiluminescence biosensor based on a 3D DNA nanomachine signal probe powered by protein–aptamer binding complex …”

Characterisation of aptamer-anchored poly(EDMA-co-GMA) monolith for high throughput affinity binding

“…Molecular interactions between aptamers and their targets are affected by physicochemical conditions of the binding environment, including ionic concentration, pH, type and characteristics of the support matrix, aptamer modifications, and temperature 18,19 . The immobilisation of aptamers to form aptasensors enhances their recycling through successive regeneration for continuous flow applications 20,21 . In recent times, the immobilisation of aptamers on macroporous monolithic matrices, with convective flow characteristics for high throughput biosensing and bioseparation applications, have emerged as a major research endeavour 22–26 .…”