Thermal Denaturation of Fresh Frozen Tissue Enhances Mass Spectrometry Detection of Peptides
Year: 2024
Type: article
Abstract: Thermal denaturation (TD), known as antigen retrieval, heats tissue samples in a buffered solution to expose protein epitopes. Thermal denaturation of formalin-fixed paraffin-embedded samples enhances on-tissue tryptic digestion, increasing peptide detection using matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI IMS). We investigated the tissue-dependent effects of TD on peptide MALDI IMS and liquid chromatography-tandem mass spectrometry signal in unfixed, frozen human colon, ovary, and pancreas tissue. In a triplicate experiment using time-of-flight, orbitrap, and Fourier-transform ion cyclotron resonance mass spectrometry platforms, we found that TD had a tissue-dependent effect on peptide signal, resulting in a (22.5%) improvement in peptide detection from the colon, a (73.3%) improvement in ovary tissue, and a (96.6%) improvement in pancreas tissue. Biochemical analysis of identified peptides shows that TD facilitates identification of hydrophobic peptides.
Source: Analytical Chemistry
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
Funders: U.S. Department of Veterans Affairs, Henry M. Jackson Foundation, Vanderbilt University, National Institutes of Health, Vanderbilt Diabetes Research and Training Center, Vanderbilt University Medical Center, National Eye Institute, National Institute of Diabetes and Digestive and Kidney Diseases, Common Fund
Awards: G1087094118, G1091621315, G1233929309, G2019238319, G2095470513, G2180887151, Biomolecular Multimodal Imaging Center: 3-Dimensional Tissue Mapping of the Human Pancreas and Eye, Flexible Hybrid Cloud Infrastructure for Seamless Integration and Use of Human Biomolecular Data and Reference Maps [1 of 5], G3606619490, G3865556588, G3989539122, G4148617410, G4654757777, Penn integrated Human Pancreas procurement and Analysis Program, G4867790825, G6766970449, G6837715458, G6845279160, G7047643307, G7260332855, G7813763335, G9000106052, G9000128600, G9000141600, G9000229981, G9000233189, G9000274329, G9000280425, Human Pancreas Analysis Program for Type 1 Diabetes - HPAP-T1D, Vanderbilt Diabetes Research Center
OpenAlex ID: W4403336294