4.5 Article

The correlation between cellular size and protein expression levels - Normalization for global protein profiling

Journal

JOURNAL OF PROTEOMICS
Volume 71, Issue 4, Pages 448-460

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.jprot.2008.06.014

Keywords

Immunohistochemistry; Protein quantification; Normalization; Expression profiling

Funding

  1. Knut and Alice Wallenberg Foundation
  2. EU

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An automated image analysis system was used for protein quantification of 1862 human proteins in 47 cancer cell lines and 12 clinical cell samples using cell microarrays and immunohistochemistry. The analysis suggests that most proteins are expressed in a cell size dependent manner, and that normalization is required for comparative protein quantification in order to correct for the inherent bias of cell size and systematic ambiguities associated with immunohistochemistry. Two reference standards were evaluated, and normalized protein expression values were found to allow for protein profiling across a panel of morphologically diverse cells, revealing putative patterns of over- and underexpression. Using this approach, proteins with stable expression as well as cell-line specific expression were identified. The results demonstrate the value of large-scale, automated proteome analysis using immunohistochemistry in revealing functional correlations and establishing methods to interpret and mine proteomic data. (C) 2008 Elsevier B.V. All rights reserved.

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