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Fluorometric visualization of mucin 1 glycans on cell surfaces based on rolling-mediated cascade amplification and CdTe quantum dots

Authors
  • Yang, XiaoTong1
  • Tang, YingYing1
  • Zhang, XiaoJing1
  • Hu, Yue1
  • Tang, Yu Ying1
  • Hu, Lin Yu1
  • Li, Su1
  • Xie, Yaochen1
  • Zhu, Dong1
  • 1 Nanjing University of Chinese Medicine, School of Pharmacy, Nanjing, Jiangsu, 210023, People’s Republic of China , Nanjing (China)
Type
Published Article
Journal
Microchimica Acta
Publisher
Springer-Verlag
Publication Date
Oct 26, 2019
Volume
186
Issue
11
Identifiers
DOI: 10.1007/s00604-019-3840-8
Source
Springer Nature
Keywords
License
Yellow

Abstract

A rolling-mediated cascade (RMC) amplification strategy is described for improved visualization of profiling glycans of mucin 1 (MUC 1) on cell surfaces. CdTe quantum dots (QDs) are used as fluorescent labels. The RMC based amplification allows even distinct glycoforms of MUC1 to be visualized on the surface of MCF-7 cell via an amplified Förster resonance energy transfer (FRET) imaging strategy that works at excitation/emission wavelengths of 345/610 nm. This is achieved by utilizing antibody against MUC1 modified with the fluorescent label 7-amino-4-methylcoumarin-3-acetic acid (AMCA) as the energy donor in FRET. The QDs (used to label surface glycans) act as acceptors. N-Azidoacetylgalactosamine-Acetylated (Ac4GalNAz) as a non-natural azido sugar, can be incorporated into the glycans of the cell surface, which can promote further labeling. The method has the advantage of only requiring a small amount of non-natural sugar to be introduced in metabolic glycan labeling since too much of an artificial sugar will interfere with the physiological functions of cells. Graphical abstractSchematic for the DNA rolling-mediated cascade (RMC)-assisted metabolic labeling of cell surface glycans by using CdTe quantum dots as labels and an intramolecular amplified FRET strategy for imaging glycans on a specific glycosylated protein, MUC1.

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