Analysis of human plasma proteome by 2DE- and 2D nanoLC-based mass spectrometry.
We compared the 2DE coupled to MALDI-TOF-MS and ESI-MS/MS analysis (2DE-MS) and the on-line 2D nanoLC, followed by nanoESI-MS/MS analysis (2DLC-MS), for the separation and identification of proteins in high abundance protein-depleted human plasma. Identification of proteins in the plasma by the two methods demonstrated that the majority of the identified protein set was unique to each method. Therefore, if a comprehensive coverage of the proteome identification is desired, it is ideal to apply both methods. The 2DE-MS method is amenable to protein spot-based quantitation, whereas the 2DLC-MS method may provide an advantage of the high throughput application.
Choi
Kyoung-Soo
KS
Department of Cellular Stress Biology, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Song
Liguo
L
Park
Young-Mee
YM
Marshall
James
J
Lund
Anders L
AL
Shion
Henry
H
Park
Eun-Mi
EM
Chae
Ho-Zoon
HZ
Park
Jong Hoon
JH
D000328:Adult; D001798:Blood Proteins / Q000032:analysis*; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001798:Blood Proteins; D020543:Proteome