Protein expression profiling of CLL B cells using replicate off-line strong cation exchange chromatography and LC-MS/MS.
In this study we use replicate 2D-LC-MS/MS analyses of crude membranes from B cells derived from a patient with chronic lymphocytic leukemia (CLL) to examine the protein expression profile of CLL B cells. Protein identifications made by replicate 2D-LC-MS/MS analysis of tryptic peptides from detergent solubilized B cell membrane proteins, as well as replicate LC-MS/MS analysis of single off-line strong cation exchange chromatography (SCX) fractions, were analyzed. We show that despite the variance in SCX, capillary LC, and the data-dependent selection of precursor ions, an overlap of 64% between proteins identified in replicate runs was achieved for this system.
Barnidge
David R
DR
Mayo Proteomics Research Center, Mayo Clinic College of Medicine, 200 1st Street SW, Rochester, MN 55905, USA. barnidge.david@mayo.edu
Tschumper
Renee C
RC
Jelinek
Diane F
DF
Muddiman
David C
DC
Kay
Neil E
NE
D015703:Antigens, CD / Q000302:isolation & purification; D001402:B-Lymphocytes / Q000378:metabolism*; D001798:Blood Proteins / Q000302:isolation & purification; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D009363:Neoplasm Proteins / Q000097:blood
D015703:Antigens, CD; D001798:Blood Proteins; D009363:Neoplasm Proteins