Magnetic microspheres modified with Ti(IV) and Nb(V) for enrichment of phosphopeptides
- 109 Downloads
Magnetic microspheres (Fe3O4) were coated with polydopamine (PDA) and loaded with the metal ions Ti(IV) and Nb(V) to give a material of type Fe3O4@PDA-Ti/Nb. It is shown to be useful for affinity chromatography and for enrichment of phosphopeptides from both standard protein solutions and real samples. For comparison, such microspheres loaded with single metal ions only (Fe3O4@PDA-Ti and Fe3O4@PDA-Nb) and their physical mixtures were also investigated under identical conditions. The binary metal ion-loaded magnetic microspheres display better enrichment efficiency than the single metal ion-loaded microspheres and their physical mixture. Both multiphosphopeptides and monophosphopeptides can be extracted. The Fe3O4@PDA-Ti/Nb microspheres exhibit ultra-high sensitivity (the lowest detection amount being 2 fmol) and selectivity at a low mass ratio such as in case of β-casein/BSA (1:1000).
KeywordsMetal ions combination Immobilized metal ion affinity chromatography MALDI-TOF mass spectrometry Polydopamine Affinity probe
This work was supported by funds provided by the Natural Science Foundation of China (Project no. 21675034), the Natural Science Foundation of Shanghai (Project no. 16ZR1402300), the Outstanding Talent Plan of Fudan University (Project no. JJF301038) and the Ministry of Science and Technology of the People’s Republic of China (Grant no. 2018ZX09J18112).
Compliance with ethical standards
The authors declare that they have no competing interests.
- 5.Di DF, Sultana R, Barone E, Perluigi M, Cini C, Mancuso C, Cai J, Pierce WM, Butterfield DA (2011) Quantitative proteomics analysis of phosphorylated proteins in the hippocampus of Alzheimer's disease subjects. J Proteome 74:1091–1103. https://doi.org/10.1016/j.jprot.2011.03.033 CrossRefGoogle Scholar
- 14.Han GH, Ye ML, Zhou HJ, Jiang XN, Feng S, Jiang XG, Tian RJ, Wan DF, Zou HF, Gu JR (2008) Large-scale phosphoproteome analysis of human liver tissue by enrichment and fractionation of phosphopeptides with strong anion exchange chromatography. Proteomics 8:1346–1361. https://doi.org/10.1002/pmic.200700884 CrossRefGoogle Scholar
- 16.Hu L, Zhou H, Li Y, Sun S, Guo L, Ye M, Tian X, Gu J, Yang S, Zou H (2009) Profiling of endogenous serum phosphorylated peptides by titanium(IV) immobilized mesoporous silica particles enrichment and MALDI-TOFMS detection. Anal Chem 81:94–104. https://doi.org/10.1021/ac801974f CrossRefGoogle Scholar
- 20.Zhao M, Deng CH, Zhang XM (2013) Synthesis of polydopamine-coated magnetic graphene for Cu2+ immobilization and application to the enrichment of low-concentration peptides for mass spectrometry analysis. ACS Appl Mater Interfaces 5:13104–13112. https://doi.org/10.1021/am4041042 CrossRefGoogle Scholar
- 22.Sun XN, Liu XD, Feng JN, Li Y, Deng CH, Duan GL (2015) Hydrophilic Nb5+-immobilized magnetic core-shell microsphere--A novel immobilized metal ion affinity chromatography material for highly selective enrichment of phosphopeptides. Anal Chim Acta 880:67–76. https://doi.org/10.1016/j.aca.2015.04.029 CrossRefGoogle Scholar
- 23.Thingholm TE, Jensen ON, Robinson PJ, Larsen MR (2008) SIMAC (sequential elution from IMAC), a phosphoproteomics strategy for the rapid separation of monophosphorylated from multiply phosphorylated peptides. Mol Cell Proteomics 7:661–671. https://doi.org/10.1074/mcp.M700362-MCP200 CrossRefGoogle Scholar
- 29.Jiang JB, Sun XN, Li Y, Deng CH, Duan GL (2018) Facile synthesis of Fe3O4@PDA core-shell microspheres functionalized with various metal ions: a systematic comparison of commonly-used metal ions for IMAC enrichment. Talanta 178:600–607. https://doi.org/10.1016/j.talanta.2017.09.071 CrossRefGoogle Scholar
- 34.Zhang L, Gan Y, Sun H, Yu B, Jin X, Zhang R, Zhang W, Zhang L (2017) Magnetic mesoporous carbon composites incorporating hydrophilic metallic nanoparticles for enrichment of phosphopeptides prior to their determination by MALDI-TOF mass spectrometry. Microchim Acta 184(2):547–555CrossRefGoogle Scholar