Pharmaceutical Sciences Faculty Publications
Document Type
Article
Publication Date
4-6-2012
Journal Title
Journal of Proteome Research
ISSN
1535-3907
Volume
11
Issue
4
First Page
2602
Last Page
2608
DOI
10.1021/pr201005t
PubMed ID
22352854
PubMed Central® ID
PMC3320745
Abstract
Formaldehyde-fixed, paraffin-embedded (FFPE) tissue repositories represent a valuable resource for the retrospective study of disease progression and response to therapy. However, the proteomic analysis of FFPE tissues has been hampered by formaldehyde-induced protein modifications, which reduce protein extraction efficiency and may lead to protein misidentification. Here, we demonstrate the use of heat augmented with high hydrostatic pressure (40,000 psi) as a novel method for the recovery of intact proteins from FFPE mouse liver. When FFPE mouse liver was extracted using heat and elevated pressure, there was a 4-fold increase in protein extraction efficiency, a 3-fold increase in the extraction of intact proteins, and up to a 30-fold increase in the number of nonredundant proteins identified by mass spectrometry, compared to matched tissue extracted with heat alone. More importantly, the number of nonredundant proteins identified in the FFPE tissue was nearly identical to that of matched fresh-frozen tissue.
Keywords
Hydrostatic pressure, liver, paraffin embedding, proteins, proteome, proteomics
Recommended Citation
Fowler, Carol B.; Waybright, Timothy J.; Veenstra, Timothy D.; O'Leary, Timothy J.; and Mason, Jeffrey T., "Pressure-Assisted Protein Extraction: A Novel Method for Recovering Proteins from Archival Tissue for Proteomic Analysis" (2012). Pharmaceutical Sciences Faculty Publications. 209.
https://digitalcommons.cedarville.edu/pharmaceutical_sciences_publications/209