Ultrasensitive Electrochemiluminescence Biosensing Platform for Detection of Multiple Types of Biomarkers toward Identical Cancer on a Single Interface

Abstract

Electrochemiluminescence (ECL) with high sensitivity and excellent controllability provides a promising approach for ultrasensitive detection of multiple biomarkers. However, the detection for multiple types of biomarkers on a single interface remains a considerable challenge owing to the functional differentiation of different types of biomarkers. Herein, we utilized "on-off-on" switching, target-induced cleavage of peptide, and TdT (terminal deoxynucleoside transferase)-mediated extension successfully constructing a novel ECL biosensor for the ultrasensitive detection of microRNA-141 (miRNA-141) and matrix metalloproteinase-2 (MMP-2). Importantly, the dual biomarkers are related with several identical cancers, which endow the biosensor with diagnostic accuracy and efficiency. In this protocol, target 1 (miRNA-141) first hybridized with probe DNA (pDNA) assembled on CdS QDs modified sensing surface. Afterward, miRNA-141 captured trigger DNA (tDNA) to generate a long ssDNA nanotail via TdT-mediated DNA polymerization. Then the forming ssDNA could capture abundant Fc-peptide-ssDNA conjugates through the hybridization reaction, the ECL intensity quenched significantly due to the efficient quenching effect of Fc to CdS QDs, realizing the ultrasensitive detection of miRNA-141 with a detection limit of 33 aM (S/N = 3). After incubated with target 2 (MMP-2) which specifically cleaved the Fc-peptide-ssDNA conjugates causing the releasing of Fc from the sensing surface, the ECL intensity had an obvious enhancement, achieving the ultrasensitive analysis of MMP-2 with a detection limit of 33 fg·mL (S/N = 3). More importantly, this biosensor also realized the monitoring of biomarkers in different cancer cells and human serum, which indicated that the biosensing system could serve as applicable tools in clinical analysis.

Modern magnetic immunoassay: Biophysical and biochemical aspects

“…As luminescent label most often are used tris (2,2′-bipyridyl) ruthenium (II). Often, such analytical systems contain avidin-biotin reagents based on the capture ELISA principle and have a quite satisfactory bioanalytical characteristic, which brings opportunities for their wide practical use (De Roeck et al, 2017;Nie et al, 2017;Wang et al, 2017b).…”

“…In particular, it becomes possible to disclaim strict requirements to the "purity" of the tested material, but also expand the range of materials that can be tested; it is possible to increase the analytical sensitivity of the analysis, to improve other bioanalytical characteristics, etc. (Tsai et al, 2007;Lin et al, 2013;Li et al, 2014;Manera et al, 2017;Nie et al, 2017).…”

Graphene-based biosensors for the detection of prostate cancer protein biomarkers: a review

“…Firstly, PSA has been demonstrated not a specific biomarker in prostate cancer early screening. As a result, detection of multiple biomarkers is crucial for precise diagnosis and prognosis of PC [130, 131]. More attention should be paid to studies on the simultaneous detection of multiple biomarkers in the future.…”

“Signal‐on” Electrogenerated Chemiluminescence Biosensing Method for the Determination of Matrix Metalloproteinase 2

“…A peptide (GPLGVRG) was reported as a substrate for the assay of MMP‐2 activity with good stability and high specificity [14]. Several peptide‐based biosensing methods for the determination of MMP‐2 have been reported, such as colorimetric [15], fluorescence [16, 17], electrochemical [18, 19] and electrogenerated chemiluminescence (electrochemiluminescence, ECL) method [20, 21]. ECL method has been studied and applied in environmental investigations, pharmaceutical analysis and clinical diagnosis with good sensitivity [22–24].…”