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A Ti/Nb-functionalized COF material based on IMAC strategy for efficient separation of phosphopeptides and phosphorylated exosomes

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Abstract

In this work, on the basis of an immobilized metal ion affinity chromatography enrichment strategy, a new kind of covalent organic framework (COF) material for enrichment of phosphorylated peptides and exosomes was successfully prepared in a facile method, and Ti4+ and Nb5+ were used as dual-functional ions (denoted as COF-S–S-COOH-Ti4+/Nb5+). With the advantage of unbiased enrichment towards phosphopeptides, COF-S–S-COOH-Ti4+/Nb5+ shows ultra-high selectivity (maximum molar ratio of β-casein: BSA is 1:20,000) and low limit of detection (0.2 fmol). In addition, the material has an excellent phosphopeptide loading capacity (100 μg/mg) and reusability (at least seven times). Furthermore, applying the material to the actual sample, 4 phosphopeptides were selectively extracted from the serum of renal carcinoma patients. At the same time, exosomes with an intact structure in the serum of renal carcinoma patients were successfully isolated rapidly using this strategy. All experiments have shown that COF-S–S-COOH-Ti4+/Nb5+ exhibits exciting potential in practical applications.

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Funding

This work is supported by the National Natural Science Foundation of China (21927805), Natural Science Foundation of Zhejiang Province (LY22B050008), Major science and technology projects in Ningbo (2020Z090), and the K. C. Wong Magna Fund in Ningbo University.

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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

    **aoya Zhang, Quanshou Feng, Zehu **e, Fuxing Xu, Yinghua Yan & Chuanfan Ding

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  1. **aoya Zhang
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  2. Quanshou Feng
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  3. Zehu **e
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  4. Fuxing Xu
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Correspondence to Fuxing Xu or Yinghua Yan.

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Ethically approved human serums 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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Zhang, X., Feng, Q., **e, Z. et al. A Ti/Nb-functionalized COF material based on IMAC strategy for efficient separation of phosphopeptides and phosphorylated exosomes. Anal Bioanal Chem 414, 7885–7895 (2022). https://doi.org/10.1007/s00216-022-04323-w

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