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.
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We sincerely appreciate the National Natural Science Foundation of China for the financial support (81573388). This work was supported by “Qing Lan Project of Jiangsu province” and “Six talent peaks project of Jiangsu Province (YY-032)”. This work was also supported by the Open Project Program of Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica (No. JKLPSE201805) and the Project of the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
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Yang, X., Tang, Y., Zhang, X. et al. Fluorometric visualization of mucin 1 glycans on cell surfaces based on rolling-mediated cascade amplification and CdTe quantum dots. Microchim Acta 186, 721 (2019). https://doi.org/10.1007/s00604-019-3840-8
- Azide polysaccharide
- Metabolic labeling
- Cancer marker
- Quantum dots
- DNA probe
- Click reaction
- Rolling-mediated cascade amplification