Analysis of glycan expression on cell surfaces by using a glassy carbon electrode modified with MnO 2 nanosheets and DNA
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Analysis of glycan expression on cell surfaces by using a glassy carbon electrode modified with MnO2 nanosheets and DNA-generated electrochemical current Kejun Feng 1 & Fangli Liao 1 & Minghui Yang 2 Received: 23 August 2019 / Accepted: 13 December 2019 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Electrochemical assay for analysis of cell surface glycan expression is reported. Mannose on human breast cancer cells (type MCF-7) is selected as the glycan model. Gold nanoparticles are modified with binding aptamer for MCF-7 cells and act as electrochemical probe. The analysis of cell surface glycan expression follows a traditional sandwich protocol. Concanavalin A that can specifically recognize mannose is immobilized onto MnO2 nanosheets modified electrode for the capture of MCF-7 cells. Then, the modified gold nanoparticles are immobilized onto the electrode via the binding between MCF-7 cell and aptamer on the gold nanoparticles. The aptamer on the gold nanoparticles reacts with molybdate. More specifically, the reaction of the phosphate backbone of aptamer with molybdate results in the formation of a redox-active molybdophosphate precipitate and generates an electrochemical current. The current intensity at 0.20 V (vs. Ag/AgCl) is recorded to test the linear range of the assay. The assay shows an obvious response to MCF-7 cells with a wide linear range from 1.0 × 103 to 1.0 × 106 cells mL−1 and a limit of detection down to 300 cells mL−1. The assay can be used to selectively monitor the change of mannose expression on cell surfaces upon the treatment with the N-glycan inhibitor. Keywords Aptamer . Concanavalin A . Gold nanoparticle . Mannose . Molybdate
Introduction Glycoproteins that generated by glycosylation are glycoconjugates of proteins. Glycoproteins are involved in nearly all biological processes and are widely found in eukaryotic cells [1–3]. Carbohydrate in glycoproteins on cell surface is participated in various physiological processes, such as cell proliferation, information transmission, growth, differentiation and progression of cancer [4, 5]. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-019-4084-3) contains supplementary material, which is available to authorized users. * Kejun Feng [email protected] 1
School of Chemistry and Materials Engineering, Huizhou University, Huizhou 516007, Guangdong, China
2
College of Chemistry and Chemical Engineering, Hunan Provincial Key Laboratory of Efficient and Clean Utilization of Manganese Resources, Central South University, Changsha 410083, China
Generally, the carbohydrate chain is long, linking numerous monosaccharides by either N-glycans or O-glycans, named according to the amino acid atom to which the carbohydrate chain is attached [6]. For example, mannose is one of the building blocks of all mammalian glycans and is the core fragment of N-glycans on the cell surface [7]. Dynamic changes of the glycan expression have been proved that related to tu
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