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 E. Kay

doi:10.1016/j.jchromb.2005.01.021

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 E. Kay

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

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 language	English (US)
Pages (from-to)	33-39
Number of pages	7
Journal	Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume	819
Issue number	1
DOIs	https://doi.org/10.1016/j.jchromb.2005.01.021
State	Published - May 5 2005

Keywords

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

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Clinical Biochemistry
Cell Biology

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10.1016/j.jchromb.2005.01.021

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Protein expression profiling of CLL B cells using replicate off-line strong cation exchange chromatography and LC-MS/MS. / Barnidge, David R.; Tschumper, Renee C.; Jelinek, Diane F. et al.
In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol. 819, No. 1, 05.05.2005, p. 33-39.

Research output: Contribution to journal › Article › peer-review

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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.",

keywords = "2D-LC-MS/MS, B cell, CLL, Expression profiling, Off-line SCX, Reproducibility",

author = "Barnidge, {David R.} and Tschumper, {Renee C.} and Jelinek, {Diane F.} and Muddiman, {David C.} and Kay, {Neil E.}",

note = "Funding Information: The authors thank Christopher Mason and Rudi Chiarito for help with data processing and bioinformatics. Supported by the National Institutes of Health grants K25 CA102148-01 (awarded to DRB), R01 CA91942 (awarded to NEK), and the Mayo Clinic College of Medicine. ",

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AU - Barnidge, David R.

AU - Tschumper, Renee C.

AU - Jelinek, Diane F.

AU - Muddiman, David C.

AU - Kay, Neil E.

N1 - Funding Information: The authors thank Christopher Mason and Rudi Chiarito for help with data processing and bioinformatics. Supported by the National Institutes of Health grants K25 CA102148-01 (awarded to DRB), R01 CA91942 (awarded to NEK), and the Mayo Clinic College of Medicine.

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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.

AB - 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.

KW - 2D-LC-MS/MS

KW - B cell

KW - CLL

KW - Expression profiling

KW - Off-line SCX

KW - Reproducibility

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

Abstract

Keywords

ASJC Scopus subject areas

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