Protein expression profiling of CLL B cells using replicate off-line strong cation exchange chromatography and LC-MS/MS

J Chromatogr B Analyt Technol Biomed Life Sci. 2005 May 5;819(1):33-9. doi: 10.1016/j.jchromb.2005.01.021.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, CD / isolation & purification
  • B-Lymphocytes / metabolism*
  • Blood Proteins / isolation & purification
  • Cell Fractionation
  • Cell Membrane / chemistry
  • Chromatography, Ion Exchange / methods*
  • Chromatography, Liquid / methods*
  • Gene Expression Profiling / methods*
  • Humans
  • Leukemia, Lymphocytic, Chronic, B-Cell / metabolism*
  • Mass Spectrometry / methods*
  • Neoplasm Proteins / blood

Substances

  • Antigens, CD
  • Blood Proteins
  • Neoplasm Proteins