title: The use of liquid phase deposition prepared phosphonate grafted silica nanoparticle-deposited capillaries in the enrichment of phosphopeptides.
abstract: In our current work, we describe how open tubular-immobilized metal-ion affinity chromatography (OT-IMAC) capillary columns connected to a solid phase microextraction (in-tube SPME) device can be used for the enrichment of phosphopeptides. A phosphonate modified silica nanoparticle (NP)-deposited capillary was prepared by liquid phase deposition (LPD), and used for the immobilization of Fe(3+), Zr(4+) or Ti(4+). The enrichment capacities of three different OT-IMAC capillary columns were compared by using tryptically digested alpha-casein as sample. The improved extraction efficiency in our technique was demonstrated by comparing to a directly modified capillary, and a comparison of phosphopeptide extraction from simple and complex samples was tested for both modes. Our results show that the NP-IMAC-Zr(4+) capillary column can be used to selectively isolate phosphopeptides from real samples, and can enrich for beta-casein phosphopeptides from concentrations as low as 1.7x10(-9) M.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan, University, Wuhan, PR China.
authors.forename: Jian-Hong
authors.initials: JH
authors.lastname: Wu
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhao
authors.forename: Ting
authors.initials: T
authors.lastname: Li
authors.forename: Cong
authors.initials: C
authors.lastname: Xu
authors.forename: Kuang
authors.initials: K
authors.lastname: Xiao
authors.forename: Yu-Qi
authors.initials: YQ
authors.lastname: Feng
authors.forename: Lin
authors.initials: L
authors.lastname: Guo
mesh_terms: D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000295:instrumentation*; D008670:Metals; D008969:Molecular Sequence Data; D053758:Nanoparticles; D063065:Organophosphonates / Q000737:chemistry*; D010748:Phosphopeptides / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D008670:Metals; D063065:Organophosphonates; D010748:Phosphopeptides; D012822:Silicon Dioxide