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Issue 12, 2016
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Competitive method-based electrochemiluminescent assay with protein–nucleotide conversion for ratio detection to efficiently monitor the drug resistance of cancer cells

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Abstract

A simple and highly-efficient approach to monitor the expression of P-glycoprotein (P-gp) in cells was urgently needed to demonstrate the drug resistance of cancer cells. Herein, a competitive method-based electrochemiluminescent (ECL) assay with a single ECL indicator was proposed for the first time to efficiently estimate the concentration ratio of two proteins. By converting the different proteins to partially coincident nucleotide sequences via a sandwich type immunoassay on magnetic beads, the concentration ratio related ECL signals could be obtained via competitive nucleotide hybridization on an electrode surface. This method could thoroughly overcome the limitations of simultaneous ECL assays via multiple ECL indicators with inevitable cross reactions. At the same time, rolling circle amplification was employed to improve the detection performances, especially the detection limit and sensitivity. With P-gp and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a model, the proposed ECL assay was successfully employed to monitor the drug resistance of cancer cells. Compared with conventional technologies, improved sensitivity and accuracy were achieved with a correlation coefficient of 0.9928 and a detection limit of 0.52%. Success in the establishment of the competitive method-based ECL assay offered an efficient strategy to demonstrate the concentration ratio of two proteins and a potential approach for detecting other proteins and nucleotide sequences, revealing a new avenue for ultrasensitive biomolecule diagnostics, especially in cell function research.

Graphical abstract: Competitive method-based electrochemiluminescent assay with protein–nucleotide conversion for ratio detection to efficiently monitor the drug resistance of cancer cells

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Publication details

The article was received on 24 Jun 2016, accepted on 04 Aug 2016 and first published on 04 Aug 2016


Article type: Edge Article
DOI: 10.1039/C6SC02801B
Citation: Chem. Sci., 2016,7, 7094-7100
  • Open access: Creative Commons BY license
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    Competitive method-based electrochemiluminescent assay with protein–nucleotide conversion for ratio detection to efficiently monitor the drug resistance of cancer cells

    W. Liang, M. Yang, Y. Zhuo, Y. Zheng, C. Xiong, Y. Chai and R. Yuan, Chem. Sci., 2016, 7, 7094
    DOI: 10.1039/C6SC02801B

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