Multiplex DNA Walking Machines for Lung Cancer-Associated miRNAs

Abstract

Biomimetic DNA walking machines have gained great success in scrutinizing the microscopic world and sensitive biosensing of disease biomarkers. Despite superb achievements, the research on DNA walking machines for simultaneous detection of multiple analytes is still rare, while the design and realization of multiplexing are considered as an important bottleneck. The multiplex detection of biomarkers can not only improve the specificity of bioassays but also avoid the squander of valuable biological specimens. Herein, we reported multiplex three-dimensional (3D) DNA walking machines based on high-resolution inductively coupled plasma mass spectrometry (HR-ICPMS) for lung cancer-associated miRNA detection. In the presence of lung cancer-associated target miRNAs (miR-21, miR-141, and miR-125b), DNA walking machines were stimulated and operated to liberate a large number of lanthanide elements (Tb, Ho, and Tm), and then the signals were collected simultaneously by HR-ICPMS. The recovery test of target miRNAs in human serum and the simultaneous monitoring experiment of three miRNAs in human lung cancer cell line (A549) and normal cell line (HBE) specimens display satisfactory analysis capabilities for complex biological samples. Thanks to the vast potential of lanthanide tags and the modular design, the proposed bioassay might flexibly detect different miRNA combinations with corresponding sets of DNA walking machines to meet the requirements of various tasks.

Simple Amplifier Coupled with a Lanthanide Labeling Strategy for Multiplexed and Specific Quantification of MicroRNAs

“…The comparison of the analytical performance of the developed method and other methods for multiplex mRNA/miRNA analysis is shown in Table S5. It can be seen that, compared with the methods based on MS for multiplex miRNA analysis, ,,,, the LODs of the developed method were lower, even lower than those methods involving the amplification technologies of DSNSA, , DNAzyme, and MNAzyme . Compared with the methods based on EDC amplification, , the developed method presented comparable LODs.…”

“…To further improve the sensitivity of the elemental labeling ICP−MS-based multiplex assay for miRNA detection, various signal amplification technologies are introduced, such as duplex-specific nuclease signal amplification (DSNSA), 16 deoxyribozyme (DNAzyme) 26 and multicomponent nucleic acid enzyme (MNAzyme) 27 amplification, and so forth. In our previous work, 27 we designed three MNAzymes that can be assembled with pairs of partzyme A/B in the presence of corresponding three target miRNAs.…”

Mass tag-encoded nanointerfaces for multiplexed mass spectrometric analysis and imaging of biomolecules

“…78 With a similar principle, Wang et al reported ICP-MS based three-dimensional (3D) DNA walking machines to evaluate lung cancer-associated miRNA-125b, miRNA-141 and miRNA-21 in lung cancer cells (A549) and normal cells (HBE), verifying the overexpression of three miRNAs in lung cancer cell lines. 91 Apart from target molecules in biofluids or cell lysates, abnormal expression of disease markers on the cell surface often reflects the pathological status of patients. 92,93 Ma et al designed bifunctional laser cleavable mass probes (LCMPs) to facilitate in situ analysis of multiple glycans, including mannose, sialic acid and N-acetylglucosamine, and to monitor dynamic alterations of glycan expression levels under the stimulation of the respective glycan inhibitor.…”

Mass Spectrometric Multiplex Detection of MicroRNA and Protein Biomarkers for Liver Cancer

“…In recent years, inductively coupled plasma mass spectrometry (ICPMS) has achieved remarkable success in the multiplex analysis of biomolecules, owing to the superior multielement analysis capability without spectral interference concomitantly. − Advantages like high sensitivity, simple operation, and quick detection offer ICPMS great potential for bioassays. − It is currently being used in the assays of nucleic acid, − protein, − enzyme, − small molecules, and single-cell assays. − Zhang et al realized dual quantification of multiple DNA assays by means of the rare element labeling strategy . Wang’s group designed a satellite structure constructed from Au NPs and up-conversion nanoparticles (UCNPs), with cancer-associated aptamers as linkers to perform a multiplexed assay of protein on exosomes .…”