Abstract
A label-free approach based on a highly reproducible and stable workflow allows for quantitative proteome analysis . Due to advantages compared to labeling methods, the label-free approach has the potential to measure unlimited samples from clinical specimen monitoring and comparing thousands of proteins. The presented label-free workflow includes a new sample preparation technique depending on automatic annotation and tissue isolation via FTIR-guided laser microdissection, in-solution digestion, LC-MS/MS analyses, data evaluation by means of Proteome Discoverer and Progenesis software, and verification of differential proteins. We successfully applied this workflow in a proteomics study analyzing human cystitis and high-grade urothelial carcinoma tissue regarding the identification of a diagnostic tissue biomarker. The differential analysis of only 1 mm2 of isolated tissue cells led to 74 significantly differentially abundant proteins.
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Acknowledgments
This work was supported by the Ministry of Innovation, Science and Research of North-Rhine Westphalia, Germany. The authors would like to thank Lidia Janota, Kristin Fuchs, Stephanie Tautges, and Birgit Zülch for their excellent technical assistance.
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Witzke, K.E., Großerueschkamp, F., Gerwert, K., Sitek, B. (2021). Application of Label-Free Proteomics for Quantitative Analysis of Urothelial Carcinoma and Cystitis Tissue. In: Marcus, K., Eisenacher, M., Sitek, B. (eds) Quantitative Methods in Proteomics. Methods in Molecular Biology, vol 2228. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1024-4_20
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DOI: https://doi.org/10.1007/978-1-0716-1024-4_20
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