The use of liquid phase deposition prepared phosphonate grafted silica nanoparticle-deposited capillaries in the enrichment of phosphopeptides.
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.
Wu
Jian-Hong
JH
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan, University, Wuhan, PR China.
Zhao
Yong
Y
Li
Ting
T
Xu
Cong
C
Xiao
Kuang
K
Feng
Yu-Qi
YQ
Guo
Lin
L
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
D008670:Metals; D063065:Organophosphonates; D010748:Phosphopeptides; D012822:Silicon Dioxide