posted on 2020-01-14, 18:04authored byDylan
M. Marchione, Ilyana Ilieva, Kyle Devins, Danielle Sharpe, Darryl J. Pappin, Benjamin A. Garcia, John P. Wilson, John B. Wojcik
Massive formalin-fixed,
paraffin-embedded (FFPE) tissue archives
exist worldwide, representing an invaluable resource for clinical
proteomics research. However, current protocols for FFPE proteomics
lack standardization, efficiency, reproducibility, and scalability.
Here we present high-yield protein extraction and recovery by direct
solubilization (HYPERsol), an optimized workflow using ultrasonication
and S-Trap sample processing that enables proteome coverage and quantification
from FFPE samples comparable to that achieved from flash-frozen tissue
(average R = 0.936). When applied to archival samples,
HYPERsol resulted in high-quality data from FFPE specimens in storage
for up to 17 years, and may enable the discovery of new immunohistochemical
markers.