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Surface functionalization modification of ultra-hydrophilic magnetic spheres with mesoporous silica for specific identification of glycopeptides in serum exosomes

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Abstract

Protein glycosylation of human serum exosomes can reveal significant physiological information, and the development of large-scale identification strategies is crucial for the in-depth investigation of the serum exosome glycoproteome. In this study, using surface functionalization techniques, an ultra-hydrophilic mesoporous silica magnetic nanosphere (denoted as Fe3O4-CG@mSiO2) was synthesized for the quick and accurate detection of glycopeptides from HRP digests. The Fe3O4-CG@mSiO2 nanospheres demonstrated outstanding enrichment capability, high sensitivity (5 amol/μL), good size exclusion effect (HRP digests/BSA proteins, 1:10,000), stable reusability (at least 10 times), and an excellent recovery rate (108.6 ± 5.5%). Additionally, after enrichment by Fe3O4-CG@mSiO2, 156 glycopeptides assigned to 64 proteins derived from human serum exosomes were successfully identified, which demonstrates that the nanospheres have great potential for the research of the large-scale serum exosome glycoproteome.

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Acknowledgements

This research was funded by Ningbo major science and technology project (2022Z133), the medical health Project of Health Department of Zhejiang Province (2021KY1045), Ningbo public welfare science and technology project (2021S104).

Ethically approved human serum samples used in this research were collected with the consent of volunteers. The study was conducted with the approval of the experimental ethics committee of Ningbo University and its affiliated hospital.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, The Affiliated Hospital of Medical School, Ningbo University, Ningbo, Zhejiang, 315020, People’s Republic of China

    Linhua Yi, Yinghua Yan, Chuan-Fan Ding & Haijiao Mao

  2. Key Laboratory of Advanced Mass Spectrometry and Molecular Analysis of Zhejiang Province, School of Material Science and Chemical Engineering, Ningbo University, Ningbo, Zhejiang, 315211, People’s Republic of China

    Baichun Wang, Quanshou Feng, Yinghua Yan & Chuan-Fan Ding

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  1. Linhua Yi
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Correspondence to Yinghua Yan or Haijiao Mao.

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Yi, L., Wang, B., Feng, Q. et al. Surface functionalization modification of ultra-hydrophilic magnetic spheres with mesoporous silica for specific identification of glycopeptides in serum exosomes. Anal Bioanal Chem 415, 1741–1749 (2023). https://doi.org/10.1007/s00216-023-04575-0

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