title: Highly selective enrichment of phosphopeptides using Zr4+-immobilized Titania nanoparticles.
abstract: It is urgent to selectively enrich trace phosphorylated proteins or peptides from complex biological sample for mass spectrometry (MS)-based phosphoproteomics. In this study, a novel immobilized metal affinity chromatography (IMAC) material with DOTA(1,4,7,10-tetraazacyclododecane N, N', N'', N'''-tetra-acetic acid) coated on the surface of TiO2nanomaterials and functionalized with the metal ion Zr4+(TiO2@DOTA-Zr) has been synthesized successfully to improve the enrichment specificity and sensitivity for phosphopeptides. TiO2nanomaterials were chosen as a support matrix, which will offer a high specific surface area to enhance the amounts of immobilized Zr4+ions and will provide an additional selectivity for the special enrichment of phosphopeptides. Also, the introduction of DOTA will endow a stronger binding interaction between the Zr4+ion and DOTA to reduce the loss of the metal ion Zr4+during sample loading and washing steps. The novel TiO2@DOTA-Zr nanohybrids exhibited higher sensitivity and selectivity for phosphopeptides from the standard protein digests, the tryptic digests of nonfat milk and human serum, showing a great selective enrichment capability toward the complicated biological samples.
authors.affiliation: College of Life Sciences, Nanjing Normal University, Nanjing 210023, China.
authors.forename: Junyong
authors.initials: J
authors.lastname: Dai
authors.affiliation: College of Zhongbei, Nanjing Normal University, Nanjing 210023, China.
authors.forename: Miaode
authors.initials: M
authors.lastname: Wang
authors.affiliation: College of Life Sciences, Nanjing Normal University, Nanjing 210023, China. Electronic address: lhl7083609@163.com.
authors.forename: Hailong
authors.initials: H
authors.lastname: Liu
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry; D006801:Humans; D008892:Milk / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry*; D010748:Phosphopeptides / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D014025:Titanium / Q000737:chemistry*; D014357:Trypsin / Q000378:metabolism; D015040:Zirconium / Q000737:chemistry*
chemical_list: D006573:Heterocyclic Compounds, 1-Ring; D010748:Phosphopeptides; C009495:titanium dioxide; C071349:1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid; D015040:Zirconium; D014025:Titanium; D014357:Trypsin
keywords: Enrichment; IMAC materials; Mass spectrometry; Phosphopeptides