| dc.description.abstract | While several proteomic studies have undertaken the challenge of analyzing formalin fixed
paraffin embedded (FFPE) tissue, few have developed optimized protocols for studying
FFPE human brain tissue. This study will examine if an improved SDS free proteomic
method applied to FFPE human brain tissue will produce a greater number of identified
proteins then assess optimized methods to produce reliable, robust and reproducible results
from this tissue. A modified guanidinium hydrochloride based extraction method was
employed and the extracted proteins were analyzed using LC-MS/MS, for shotgun
proteomics. The FFPE human brain tissue samples were acquired from archived specimens
at the Griffith University School of Anatomy. By analyzing tissue samples from human
FFPE brain, we evaluated conditions used for tissue lysis, digestion, and the effects of the
fixation time and storage duration on the number of proteins obtained during shotgun
proteomic analysis on FFPE tissue. Comparing the identified proteins and experimental
procedures to the existing FFPE tissue studies in the literature, the limitations due to protein
quality were identified, optimized processes proposed, and future studies including multiomic analysis of the human brain, were outlined. | en_US |
