Solid-phase extraction and elution on diamond (SPEED): a fast and general platform for proteome analysis with mass spectrometry.
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.
Chen
Wei-Hao
WH
Institute of Atomic and Molecular Sciences, Academia Sinica, P.O. Box 23-116, Taipei 106, Taiwan, ROC.
Lee
Sheng-Chung
SC
Sabu
Sahadevan
S
Fang
Huei-Chun
HC
Chung
Shu-Chien
SC
Han
Chau-Chung
CC
Chang
Huan-Cheng
HC
D000595:Amino Acid Sequence; D018130:Diamond; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004867:Equipment Design; D006801:Humans; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000379:methods; D011506:Proteins / Q000032:analysis / Q000737:chemistry; D011507:Proteinuria; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D004220:Disulfides; D011506:Proteins; D018130:Diamond