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Additions to the Human Plasma Proteome via a Tandem MARS Depletion iTRAQ-Based Workflow

Cao, Zhiyun and Yende, Sachin and Kellum, John A. and Robinson, Renã A. S. (2013) Additions to the Human Plasma Proteome via a Tandem MARS Depletion iTRAQ-Based Workflow. International Journal of Proteomics, 2013. pp. 1-8. ISSN 2090-2166

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Robust platforms for determining differentially expressed proteins in biomarker and discovery studies using human plasma are of great interest. While increased depth in proteome coverage is desirable, it is associated with costs of experimental time due to necessary sample fractionation. We evaluated a robust quantitative proteomics workflow for its ability (1) to provide increased depth in plasma proteome coverage and (2) to give statistical insight useful for establishing differentially expressed plasma proteins. The workflow involves dual-stage immunodepletion on a multiple affinity removal system (MARS) column, iTRAQ tagging, offline strong-cation exchange chromatography, and liquid chromatography tandem mass spectrometry (LC-MS/MS). Independent workflow experiments were performed in triplicate on four plasma samples tagged with iTRAQ 4-plex reagents. After stringent criteria were applied to database searched results, 689 proteins with at least two spectral counts (SC) were identified. Depth in proteome coverage was assessed by comparison to the 2010 Human Plasma Proteome Reference Database in which our studies reveal 399 additional proteins which have not been previously reported. Additionally, we report on the technical variation of this quantitative workflow which ranges from ±11 to 30%.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Cao, Zhiyun
Yende, Sachin
Kellum, John A.
Robinson, Renã A.
Date: 2013
Date Type: Publication
Journal or Publication Title: International Journal of Proteomics
Volume: 2013
Publisher: Hindawi Publishing Corporation
Page Range: pp. 1-8
DOI or Unique Handle: 10.1155/2013/654356
Schools and Programs: Dietrich School of Arts and Sciences > Chemistry
Refereed: Yes
ISSN: 2090-2166
Official URL:
Article Type: Research Article
Date Deposited: 04 May 2020 17:37
Last Modified: 04 May 2020 17:37


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