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A novel hydrophilic polymer-coated magnetic nanomaterial based on the HILIC strategy for fast separation of glycopeptides and glycosylated exosomes

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Abstract

Novel hydrophilic poly(N, N-methylenebisacrylamide/1,2-epoxy-5-hexene) coated magnetic nanospheres functionalized with 2-aminopurine (denoted as Fe3O4@poly(MBA/EH)@2AP) for enriching glycopeptides and glycosylated exosomes were successfully obtained using a simple and green method on the basis of the HILIC (hydrophilic interaction liquid chromatography) enrichment strategy. The high density of polar groups endows the material with amazing hydrophilicity, enabling the nanomaterial to successfully capture glycopeptides and glycosylated exosomes within 1 min. Meanwhile, the materials demonstrated great sensitivity (0.01 fmol/μL), good loading capability (125 μg/mg), high selectivity (BSA:HRP = 1000:1), and repeatability (more than 10 times). Besides, the material was applied in the analysis of bio-samples, a total of 290 glycosylated peptides and 184 glycosylation sites mapping to 185 glycoproteins were identified in the serum of uremic patients. Besides, 42 glycopeptides were enriched from the saliva of healthy people. At the same time, it was verified by TEM and western blot that the complete glycosylated exosomes were successfully captured from the serum of the uremic patients. All experiments have demonstrated that Fe3O4@poly(MBA/EH)@2AP has a promising future in practical applications.

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Funding

This research is financed by the Natural Science Foundation of Zhejiang Province (LY22B050008).

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Authors and Affiliations

  1. Key Laboratory of Advanced Mass Spectrometry and Molecular Analysis of Zhejiang Province, Institute of Mass Spectrometry, School of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, Zhejiang, China

    Xiaoya Zhang, Shuweng Hua, Quanshou Feng, Chuan-Fan Ding & Yinghua Yan

  2. Li Huili East Hospital of Ningbo Medical Center, Ningbo, China

    Yongyao Wu

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  1. Xiaoya Zhang
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  2. Shuweng Hua
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  3. Quanshou Feng
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Correspondence to Chuan-Fan Ding or Yinghua Yan.

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Samples of human serum were obtained from the Affiliated Hospital of Ningbo University with the approval of its Ethics Committee (KS20227002).

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The ethically approved human serum and saliva used in this study were collected with the consent of the volunteers. This study was approved by the experimental ethics committee of Ningbo University and its affiliated hospital.

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Zhang, X., Hua, S., Feng, Q. et al. A novel hydrophilic polymer-coated magnetic nanomaterial based on the HILIC strategy for fast separation of glycopeptides and glycosylated exosomes. Anal Bioanal Chem 415, 5755–5767 (2023). https://doi.org/10.1007/s00216-023-04857-7

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