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Anal Chim Acta. 2018 Dec 18;1039:108-115. doi: 10.1016/j.aca.2018.07.040. Epub 2018 Jul 19.

Lectin-mediated in situ rolling circle amplification on exosomes for probing cancer-related glycan pattern.

Author information

1
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, PR China.
2
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, PR China. Electronic address: dinglin@nju.edu.cn.
3
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, PR China. Electronic address: wujie@nju.edu.cn.

Abstract

Exosomal surface glycans play important roles in microvesicle protein sorting and exosome-cell interactions, and also provide promising biomarkers for various diseases. However, in situ detection techniques for exosomal glycans are largely lacking. In this work, an exosomal array is fabricated for probing cancer-related exosomal glycan signatures by lectin recognition-mediated in situ rolling circle assembly of fluorophore-labeled DNA. Different from commonly used lectin array, the proposed strategy enables the direct and amplified conversion of glycan recognition signals to fluorescence detection signals. Focusing on tumor-associated glycans including sialic acids, fucose and truncated O-glycans, the method has been used not only to compare glycan patterns between exosomes with different origins, but also to reveal the specific exosomal glycan characteristics compared to their parent cells. The limits of detection were identified to be 5.4 × 106 and 1.3 × 106 particles mL-1 for HeLa and PANC-1 exosomes, respectively. The dynamic ranges were 4.7 × 105 to 4.7 × 108, 4.7 × 108 to 4.7 × 109 for HeLa exosomes, and 4.7 × 105 to 1.2 × 109, 1.2 × 109 to 4.7 × 109 particles mL-1 for PANC-1 exosomes. The remodeling of exosomal glycans can also be monitored as demonstrated on the cleavage of sialic acids under sialidase treatment. It could be anticipated that this strategy would become a powerful tool for development of exosome-based glyco-biomarkers and elucidation of biological significance of exosomal glycans.

KEYWORDS:

Array; Exosome; Glycan; In situ; Lectin; Rolling circle amplification

PMID:
30322541
DOI:
10.1016/j.aca.2018.07.040

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