Targeted proteomic approach for quantification of collagen type I and type III in formalin-fixed paraffin-embedded tissue.

Autor: Rosin NL; Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada., Winstone TML; Department of Pathology and Laboratory Medicine, University of Calgary, 3535 Research Rd NW, Room 1E-415, Calgary, AB, T2I 2K8, Canada., Kelley M; Department of Pathology and Laboratory Medicine, University of Calgary, 3535 Research Rd NW, Room 1E-415, Calgary, AB, T2I 2K8, Canada., Biernaskie J; Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.; Department of Surgery, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.; Hotchkiss Brain Institute, University of Calgary, Calgary, AB, Canada.; Alberta Children's Hospital Research Institute, University of Calgary, Calgary, AB, Canada., Dufour A; Department of Physiology and Pharmacology, University of Calgary, Calgary, AB, Canada.; McCaig Institute for Bone and Joint Health, University of Calgary, Calgary, AB, Canada., Orton DJ; Department of Pathology and Laboratory Medicine, University of Calgary, 3535 Research Rd NW, Room 1E-415, Calgary, AB, T2I 2K8, Canada. dennis.orton@ucalgary.ca.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Aug 01; Vol. 14 (1), pp. 17769. Date of Electronic Publication: 2024 Aug 01.
DOI: 10.1038/s41598-024-68377-9
Abstrakt: Collagen is the most abundant protein in mammals and a major structural component of the extracellular matrix (ECM). Changes to ECM composition occur as a result of numerous physiological and pathophysiological causes, and a common means to evaluate these changes is the collagen 3 (Col3) to collagen 1 (Col1) ratio. Current methods to measure the Col3/1 ratio suffer from a lack of specificity and often under- or over-estimate collagen composition and quantity. This manuscript presents a targeted liquid chromatography tandem mass spectrometry (LC-MS/MS) method for quantification of Col3 and Col1 in FFPE tissues. Using surrogate peptides to generate calibration curves, Col3 and Col1 are readily quantified in FFPE tissue sections with high accuracy and precision. The method is applied to several tissue types from both human and reindeer sources, demonstrating its generalizability. In addition, the targeted LC-MS/MS method permits quantitation of the hydroxyprolinated form of Col3, which has significant implications for understanding not only the quantity of Col3 in tissue, but also understanding of the pathophysiology underlying many causes of ECM changes. This manuscript presents a straightforward, accurate, precise, and generalizable method for quantifying the Col3/1 ratio in a variety of tissue types and organisms.
(© 2024. The Author(s).)
Databáze: MEDLINE
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