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Nano Research

, Volume 12, Issue 6, pp 1473–1481 | Cite as

Reproducible large-scale synthesis of surface silanized nanoparticles as an enabling nanoproteomics platform: Enrichment of the human heart phosphoproteome

  • David S. Roberts
  • Bifan Chen
  • Timothy N. Tiambeng
  • Zhijie Wu
  • Ying GeEmail author
  • Song JinEmail author
Research Article
  • 173 Downloads

Abstract

A reproducible synthetic strategy was developed for facile large-scale (200 mg) synthesis of surface silanized magnetite (Fe3O4) nanoparticles (NPs) for biological applications. After further coupling a phosphate-specific affinity ligand, these functionalized magnetic NPs were used for the highly specific enrichment of phosphoproteins from a complex biological mixture. Moreover, correlating the surface silane density of the silanized magnetite NPs to their resultant enrichment performance established a simple and reliable quality assurance control to ensure reproducible synthesis of these NPs routinely in large scale and optimal phosphoprotein enrichment performance from batch-to-batch. Furthermore, by successful exploitation of a top-down phosphoproteomics strategy that integrates this high throughput nanoproteomics platform with online liquid chromatography (LC) and tandem mass spectrometry (MS/MS), we were able to specifically enrich, identify, and characterize endogenous phosphoproteins from highly complex human cardiac tissue homogenate. This nanoproteomics platform possesses a unique combination of scalability, specificity, reproducibility, and efficiency for the capture and enrichment of low abundance proteins in general, thereby enabling downstream proteomics applications.

Keywords

nanoparticles nanoproteomics surface functionalization large-scale phosphoprotein enrichment top-down proteomics mass spectrometry 

Notes

Acknowledgements

The financial support for this project is provided by NIH R01 GM117058 (to S. J. and Y. G.). Moreover, Y. G. would like to acknowledge the NIH R01 GM125085 and S10 OD018475. T. N. T. would like to acknowledge support from the NIH Chemistry-Biology Interface Training Program NIH T32GM008505.

Supplementary material

12274_2019_2418_MOESM1_ESM.pdf (3.7 mb)
Reproducible large-scale synthesis of surface silanized nanoparticles as an enabling nanoproteomics platform: Enrichment of the human heart phosphoproteome

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Copyright information

© Tsinghua University Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of ChemistryUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Department of Cell and Regenerative BiologyUniversity of Wisconsin-MadisonMadisonUSA

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