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

• Published in: Nano research Vol. 12; no. 6; pp. 1473 - 1481
• Main Authors: Roberts, David S., Chen, Bifan, Tiambeng, Timothy N., Wu, Zhijie, Ge, Ying, Jin, Song
• Format: Journal Article
• Language: English
• Published: Beijing Tsinghua University Press 01-06-2019; Springer Nature B.V
• Subjects: Atomic/Molecular Structure and Spectra; Biomedicine; Biotechnology; Chemistry and Materials Science; Condensed Matter Physics; Enrichment; Heart; Iron oxides; Liquid chromatography; Magnetite; Mass spectrometry; Mass spectroscopy; Materials Science; Nanoparticles; Nanotechnology; Phosphoproteins; Proteins; Proteomics; Quality assurance; Reproducibility; Research Article; Synthesis; nanoparticles; top-down proteomics; surface functionalization; nanoproteomics; mass spectrometry; phosphoprotein enrichment; large-scale
• ISSN: 1998-0124; 1998-0000
• DOI: 10.1007/s12274-019-2418-4

A reproducible synthetic strategy was developed for facile large-scale (200 mg) synthesis of surface silanized magnetite (Fe
3
O
4
) 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.