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In situ digestion-assisted multi-template imprinted nanoparticles for efficient analysis of protein phosphorylation

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Abstract

A new general approach called in situ digestion-assisted multi-template imprinting is proposed for preparation of phospho-specific molecularly imprinted nanoparticles. Through the novel templating strategy and controllable imprinting process, imprinted nanoparticles specific to the intact phosphoprotein and its phosphopeptides were synthesized. The prepared imprinted nanoparticles exhibited excellent specificity (cross reactivity < 10%), high affinity (10−6 M), high efficiency (47.5%), and good generality (both intact phosphoprotein and phosphopeptides). We also realized the fine tuning of the recognition at peptide level of the imprinted nanoparticles by adjusting the imprinting time. Based on the selective enrichment of the imprinted nanoparticles, the MS identification of both the intact phosphoprotein (Tau) and phosphopeptides (angiotensin II and peptides of Tau) in real complex samples could be achieved. Therefore, we believe that the in situ digestion-assisted multi-template imprinting strategy holds promising future in both phosphorylation analysis and proteomics applications.

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Funding

The authors received financial support of the 512 Talent Cultivation Plan of Bengbu Medical College (by51202305), the Natural Science Research Project of Bengbu Medical college (2022byfy003), and the Natural Science Research Project of Anhui Educational Committee (2022AH051537, 2022AH051424, 2023AH030091).

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Authors and Affiliations

  1. Department of Chemistry, Bengbu Medical University, 2600 Donghai Avenue, Bengbu, 233030, China

    Jie Wang, Hui Zhang, Yang Chen & Zijun Bie

  2. School of Pharmacy, Bengbu Medical University, 2600 Donghai Avenue, Bengbu, 233000, China

    Xiuling Zhao, Yang Chen & Zijun Bie

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  1. Jie Wang
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Wang, J., Zhao, X., Zhang, H. et al. In situ digestion-assisted multi-template imprinted nanoparticles for efficient analysis of protein phosphorylation. Microchim Acta 190, 490 (2023). https://doi.org/10.1007/s00604-023-06081-7

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