Solid-Phase Extraction and Elution on Diamond (Speed): A Fast and General Platform for Proteome Analysis with Mass Spectrometry
Resource
ANALYTICAL CHEMISTRY v.78 n.12 pp.4228-4234
Journal
ANALYTICAL CHEMISTRY
Journal Volume
v.78
Journal Issue
n.12
Pages
4228-4234
Date Issued
2006
Date
2006
Author(s)
LEE, SHENG-CHUNG
Abstract
This paper presents a new solid-phase extraction and elution platform based on surface-functionalized diamond nanocrystallites. Compared with conventional methods, the platform facilitates purification and concentration of intact proteins and their enzymatic digests for ensuing sodium dodecyl sulfate-polyacrylamide gel electrophoresis or matrix- assisted laser desorption/ionization mass spectrometry analysis without prior removal of the adsorbent . One-pot work flow involving reduction of disulfide bonds, protection of free cysteine residues, washing off residual chemicals, and proteolytic digestion of adsorbed proteins can be performed directly on the particles. The utility and versatility of this protein workup platform were demonstrated with liquid chromatography- electrospray ionization tandem mass spectrometry in proteome analysis of human urine. The proteome analysis of each urine sample can be completed in 8 h.
Subjects
2-DIMENSIONAL ELECTROPHORESIS GELS
CEREBROSPINAL-FLUID
HUMAN URINE
FUNCTIONAL PROTEOMICS
AFFINITY CAPTURE, HUMAN PLASMA
PROTEINS