Abstract
In this study, titanium (IV)-immobilized magnetic nanoparticles (Ti4+-PTL-MNPs) were firstly synthesized via a one-step aqueous self-assembly of lysozyme nanofilms for efficient phosphopeptide enrichment. Under physiological conditions, lysozymes readily self-organized into phase-transitioned lysozyme (PTL) nanofilms on Fe3O4@SiO2 and Fe3O4@C MNP surfaces with abundant functional groups, including –NH2, –COOH, –OH, and –SH, which can be used as multiple linkers to efficiently chelate Ti4+. The obtained Ti4+-PTL-MNPs possessed high sensitivity of 0.01 fmol μL−1 and remarkable selectivity even at a mass ratio of β-casein to BSA as low as 1:400 for phosphopeptide enrichment. Furthermore, the synthesized Ti4+-PTL-MNPs can also selectively identify low-abundance phosphopeptides from extremely complicated human serum samples and their rapid separation, good reproducibility, and excellent recovery were also proven. This one-step self-assembly of PTL nanofilms facilitated the facile and efficient surface functionalization of various nanoparticles for proteomes/peptidomes.
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Acknowledgements
This work was supported by the Natural Science Foundation Project of Shaanxi Province (No. 2020JZ-24), the National Key R&D Program of China (2019YFB2103000), and the Fundamental Research Funds for the Central Universities (GK201801006).
Funding
National Key R&D Program of China, 2019 YFB2103000, Fundamental Research Funds for the Central Universities, GK201801006, Natural Science Foundation Project of Shaanxi Province, No. 2020JZ-24
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Jianru Li & Nan Li: Conceptualization, Methodology, Investigation, Validation, Writing—original draft, Writing—review & editing. Yawen Hou: Investigation, Validation. Miao Fan & Yuxiu Zhang & Qiqi Zhang: Resources. Fuquan Dang: Supervision, Funding acquisition.
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Human serum was obtained from Shaanxi Normal University Hospital. All the experiments were permitted by Human Research Ethics Board of Shaanxi Normal University (NO. 20150323). All participants provided written informed consent.
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Li, J., Li, N., Hou, Y. et al. Facile fabrication of Ti4+-immobilized magnetic nanoparticles by phase-transitioned lysozyme nanofilms for enrichment of phosphopeptides. Anal Bioanal Chem 416, 1657–1665 (2024). https://doi.org/10.1007/s00216-024-05170-7
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DOI: https://doi.org/10.1007/s00216-024-05170-7