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

David R. Barnidge, Renee C. Tschumper, Diane F Jelinek, David C. Muddiman, Neil Elliot Kay

Research output: Contribution to journalArticle

12 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)33-39
Number of pages7
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume819
Issue number1
DOIs
StatePublished - May 5 2005

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B-Cell Chronic Lymphocytic Leukemia
Chromatography
Cations
Cells
B-Lymphocytes
Proteins
Detergents
Membrane Proteins
Ions
Membranes
Peptides

Keywords

  • 2D-LC-MS/MS
  • B cell
  • CLL
  • Expression profiling
  • Off-line SCX
  • Reproducibility

ASJC Scopus subject areas

  • Biochemistry

Cite this

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title = "Protein expression profiling of CLL B cells using replicate off-line strong cation exchange chromatography and LC-MS/MS",
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.",
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T1 - Protein expression profiling of CLL B cells using replicate off-line strong cation exchange chromatography and LC-MS/MS

AU - Barnidge, David R.

AU - Tschumper, Renee C.

AU - Jelinek, Diane F

AU - Muddiman, David C.

AU - Kay, Neil Elliot

PY - 2005/5/5

Y1 - 2005/5/5

N2 - 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.

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KW - 2D-LC-MS/MS

KW - B cell

KW - CLL

KW - Expression profiling

KW - Off-line SCX

KW - Reproducibility

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