title: Immobilized metal ion affinity chromatography ZipTip pipette tip with polydopamine modification and Ti⁴⁺ immobilization for selective enrichment and isolation of phosphopeptides.
abstract: As an effective tool in protein analysis, mass spectroscopy (MS) has been widely used in identifying protein phosphorylation and phosphorylation sites. Because of the low abundance of phosphopeptides in protein digestion, isolation and enrichment of phosphopeptides prior to MS analysis is important for efficient phosphopeptides identification. In this work, we initially immobilized titanium ions on polydopamine (PDA)-modified ZipTip pipette tips (denoted as IMAC ZipTip pipette tip) for simple and quick enrichment of phosphopeptides. The preparation process of the novel ZipTip pipette tips is fast and economic since it only contains two simple steps both with mild conditions. The ability of modified ZipTip pipette tips for identifying phosphopeptides in complex biological samples was investigated. The unique ZipTip pipette tip not only exhibited superior ability in selectively capturing phosphopeptides from large amount of non-phosphopeptides, but also remarkably shortened the MS preparation and analysis time, making it an easy-to-use and efficient tool in phosphoproteome research.
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Chenyi
authors.initials: C
authors.lastname: Shi
authors.affiliation: Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, China.. Electronic address: chdeng@fudan.edu.cn.
authors.forename: Chunhui
authors.initials: C
authors.lastname: Deng
mesh_terms: D002364:Caseins / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D007211:Indoles / Q000737:chemistry*; D010748:Phosphopeptides / Q000097:blood / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D014025:Titanium / Q000737:chemistry*
chemical_list: D002364:Caseins; D007211:Indoles; D010748:Phosphopeptides; D011108:Polymers; C568283:polydopamine; D012710:Serum Albumin, Bovine; D014025:Titanium
keywords: Immobilized metal ion affinity chromatography technique; Mass spectrometry; Phosphopeptides identification; ZipTip pipette tip