Evaluation of protein purification techniques and effects of storage duration on lc-ms/ms analysis of archived ffpe human crc tissues
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Date
2021
Journal Title
Journal ISSN
Volume Title
Publisher
Frontiers Media
Abstract
To elucidate cancer pathogenesis and its mechanisms at the molecular level, the collecting
and characterization of large individual patient tissue cohorts are required. Since most
pathology institutes routinely preserve biopsy tissues by standardized methods of formalin
fixation and paraffin embedment, these archived FFPE tissues are important collections of
pathology material that include patient metadata, such as medical history and treatments.
FFPE blocks can be stored under ambient conditions for decades, while retaining cellular
morphology, due to modifications induced by formalin. However, the effect of long-term
storage, at resource-limited institutions in developing countries, on extractable protein
quantity/quality has not yet been investigated. In addition, the optimal sample preparation
techniques required for accurate and reproducible results from label-free LC-MS/MS
analysis across block ages remains unclear. This study investigated protein extraction
efficiency of 1, 5, and 10-year old human colorectal carcinoma resection tissue and
assessed three different gel-free protein purification methods for label-free LC-MS/MS
analysis. A sample size of n 17 patients per experimental group (with experiment power
0.7 and α 0.05, resulting in 70% confidence level) was selected.
Description
Keywords
Protein, Cancer, Patient tissue, Paraffin
Citation
Rossouw, S. C. et al. (2021). Evaluation of protein purification techniques and effects of storage duration on lc-ms/ms analysis of archived ffpe human crc tissues. Pathology and Oncology Research, 27. https://doi.org/10.3389/pore.2021.622855