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.
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The authors thank the support of this work by the Program for Innovative Research Team of Huizhou University and the Hunan Provincial Science and Technology Plan Project, China (no. 2016TP1007).
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Feng, K., Liao, F. & Yang, M. Analysis of glycan expression on cell surfaces by using a glassy carbon electrode modified with MnO2 nanosheets and DNA-generated electrochemical current. Microchim Acta 187, 148 (2020). https://doi.org/10.1007/s00604-019-4084-3
- Concanavalin A
- Gold nanoparticle