Document Detail


Improved protein extraction and protein identification from archival formalin-fixed paraffin-embedded human aortas.
MedLine Citation:
PMID:  23339088     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: Evaluate combination of heat and elevated pressure to enhance protein extraction and quality of formalin-fixed (FF), and FF paraffin-embedded (FFPE) aorta for proteomics.
EXPERIMENT DESIGN: Proteins were extracted from fresh frozen aorta at room temperature (RT). FF and FFPE aortas (3 months and 15 years) were extracted at RT, heat alone, or a combination of heat and high pressure. Protein yields were compared, and digested peptides from the extracts were analyzed with MS.
RESULTS: Combined heat and elevated pressure increased protein yield from human FF or FFPE aorta compared to matched tissues with heat alone (1.5-fold) or at RT (8.3-fold), resulting in more proteins identified and with more sequence coverage. The length of storage did adversely affect the quality of proteins from FF tissue. For long-term storage, aorta was preserved better with FFPE than FF alone. Periostin and MGF-E8 were demonstrated suitable for MRM assays from FFPE aorta.
CONCLUSIONS AND CLINICAL RELEVANCE: Combination of heat and high pressure is an effective method to extract proteins from FFPE aorta for downstream proteomics. This method opens the possibility for use of archival and often rare FFPE aortas and possibly other tissues available to proteomics for biomarker discovery and quantification.
Authors:
Zongming Fu; Kun Yan; Avraham Rosenberg; Zhicheng Jin; Barbara Crain; Grace Athas; Richard S Vander Heide; Timothy Howard; Allen D Everett; David Herrington; Jennifer E Van Eyk
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Proteomics. Clinical applications     Volume:  7     ISSN:  1862-8354     ISO Abbreviation:  Proteomics Clin Appl     Publication Date:  2013 Apr 
Date Detail:
Created Date:  2013-04-15     Completed Date:  2013-12-18     Revised Date:  2014-06-06    
Medline Journal Info:
Nlm Unique ID:  101298608     Medline TA:  Proteomics Clin Appl     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  217-24     Citation Subset:  IM    
Copyright Information:
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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MeSH Terms
Descriptor/Qualifier:
Aorta / metabolism*,  pathology
Biological Markers / chemistry
Fixatives / chemistry*
Formaldehyde / chemistry*
Humans
Paraffin / chemistry*
Paraffin Embedding / methods*
Proteins / isolation & purification*
Proteomics
Tissue Fixation / methods
Grant Support
ID/Acronym/Agency:
5RC1HL100021-02/HL/NHLBI NIH HHS; R01 HL111362/HL/NHLBI NIH HHS; RC1 HL100021/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Biological Markers; 0/Fixatives; 0/Proteins; 1HG84L3525/Formaldehyde; 8002-74-2/Paraffin

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