Application of open tubular capillary columns coated with zirconium phosphonate for enrichment of phosphopeptides.
A new approach utilizing open tubular capillary columns coated with zirconium phosphonate (ZrP-OTCC) for enrichment of phosphopeptides is described. The experimental conditions: interior diameter, length of capillary and flow rate was optimized using tryptic digest of alpha-casein (a phosphoprotein) as a model sample. The ZrP-OTCC was demonstrated to tolerate urea, sodium dodecyl sulphate (SDS), and NaCl. Further experimental results show that the ZrP-OTCC can trap the phosphopeptides even at the concentration of alpha-casein as low as 10(-8)M. This column has also been successfully coupled online with nano-liquid chromatography for enrichment and then separation of phosphopeptides from a complex sample, and finally analyzed the phosphopeptides by mass spectrometry (MS).
Xue
Yanfeng
Y
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, No. 33 Life Science Park Road, Changping District, Beijing 102206, PR China.
Wei
Junying
J
Han
Huanhuan
H
Zhao
Liyan
L
Cao
Dong
D
Wang
Jinglan
J
Yang
Xiaoming
X
Zhang
Yangjun
Y
Qian
Xiaohong
X
D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods*; D010748:Phosphopeptides / Q000302:isolation & purification*; D015040:Zirconium / Q000737:chemistry*
D010748:Phosphopeptides; C027006:zirconium phosphate; D015040:Zirconium