Journal of Proteomics & Bioinformatics
Open Access
ISSN: 0974-276X
ISSN: 0974-276X
Jeffrey T Mason
Tanzania
Research Article
Improving the Proteomic Analysis of Archival Tissue by Using Pressure-
Assisted Protein Extraction: A Mechanistic Approach
Author(s): Carol B Fowler, Timothy J O'Leary and Jeffrey T MasonCarol B Fowler, Timothy J O'Leary and Jeffrey T Mason
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 tissue. Our laboratory has taken a mechanistic approach to developing improved protein extraction protocols, by first studying the reactions of formaldehyde with proteins and ways to reverse these reactions, then applying this approach to a model system called a “tissue surrogate”, which is a gel formed by treating high concentrations of cytoplasmic proteins.. View More»
DOI:
10.4172/jpb.1000315