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Application of 2-D DIGE to formalin-fixed, paraffin-embedded tissues

Tanca, Alessandro and Pagnozzi, Daniela and Falchi, Giovanni and Tonelli, Roberto and Rocca, Stefano and Roggio, Tonina and Uzzau, Sergio and Addis, Maria Filippa (2011) Application of 2-D DIGE to formalin-fixed, paraffin-embedded tissues. Proteomics, Vol. 11 (5), p. 1005-1011. eISSN 1615-9861. Article.

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DOI: 10.1002/pmic.201000353

Abstract

The ability to investigate the proteome of formalin-fixed, paraffin-embedded (FFPE) tissues can be considered a major recent achievement in the field of clinical proteomics. However, gel-based approaches to the investigation of FFPE tissue proteomes have lagged behind, mainly because of insufficient quality of full-length protein extracts. Here, the 2-D DIGE technology was investigated for applicability to FFPE proteins, for internal reproducibility among replicate FFPE extracts, and for comparability between FFPE and fresh-frozen tissue profiles. The 2-D DIGE patterns obtained upon labeling and electrophoresis of replicate FFPE tissue extracts were highly reproducible, with satisfactory resolution and complexity. Moreover, the implementation of DIGE enabled to highlight and characterize the consistent differences found in the FFPE profiles compared with fresh-frozen profiles, represented by an acidic shift, directly correlated to the protein pI value, and by a reduction in spot signal intensity, directly correlated to molecular weight and percentage of lysine residues. Being constantly and reproducibly present in all FFPE tissue extract replicates at similar extents, these modifications do not appear to hinder the comparative analysis of FFPE tissue extracts by 2-D DIGE, opening the way to its application for the differential proteomic investigation of archival tissue repositories.

Item Type:Article
ID Code:5453
Status:Published
Refereed:Yes
Uncontrolled Keywords:2-D DIGE, acidic shift, archived tissues, formalin-fixed, paraffin embedded, lysine, technology
Subjects:Area 06 - Scienze mediche > MED/07 Microbiologia e microbiologia clinica
Area 07 - Scienze agrarie e veterinarie > VET/03 Patologia generale e anatomia patologica veterinaria
Divisions:001 Università di Sassari > 01 Dipartimenti > Patologia e clinica veterinaria
001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
001 Università di Sassari > 01 Dipartimenti > Protezione delle piante
002 Altri enti e centri di ricerca del Nord Sardegna > Porto Conte Ricerche, Alghero
002 Altri enti e centri di ricerca del Nord Sardegna > Biosistema s.c.r.l.-Centro di competenza e trasferimento tecnologico biosistema, Sassari
Publisher:Wiley
eISSN:1615-9861
Deposited On:10 Feb 2011 09:25

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