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Liquid chromatography-tandem mass spectrometry analysis of peptides separated from the insoluble matrix by in-sample tryptic protein digestion for rapid discrimination of various induced pathological states of human bone models in oral surgery
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SYSNO ASEP 0563451 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Liquid chromatography-tandem mass spectrometry analysis of peptides separated from the insoluble matrix by in-sample tryptic protein digestion for rapid discrimination of various induced pathological states of human bone models in oral surgery Author(s) Michalus, I. (CZ)
Van Nguyen, T. (CZ)
Viktorová, J. (CZ)
Cejnar, P. (CZ)
Šantrůček, J. (CZ)
Kučková, Š. (CZ)
Sázelová, Petra (UOCHB-X) RID, ORCID
Kašička, Václav (UOCHB-X) RID, ORCID
Hynek, R. (CZ)Source Title Journal of Separation Science. - : Wiley - ISSN 1615-9306
Roč. 45, č. 24 (2022), s. 4388-4396Number of pages 9 s. Language eng - English Country DE - Germany Keywords liquid chromatography ; mass spectrometry ; protein digestion ; cancer ; inflammation ; in-sample protein digestion OECD category Analytical chemistry Method of publishing Limited access Institutional support UOCHB-X - RVO:61388963 UT WOS 000871905200001 EID SCOPUS 85140450325 DOI 10.1002/jssc.202200694 Annotation For the understanding of pathological states of bone tissues in oral surgery, it would be desirable to have the possibility to simulate these processes on bone cell models in vitro. These cultures, similarly to bone tissues, contain numerous proteins entrapped in the insoluble matrix. The major goal of this study was to verify whether a method based on direct in-matrix protein digestion could be suitable for the discrimination between different induced pathological states of bone cell models cultivated in vitro. Using in-sample specific protein digestion with trypsin followed by liquid chromatography-tandem mass spectrometry analysis of released peptides, 446 proteins (in average per sample) were identified in a bone cell in vitro model with induced cancer, 440 proteins were found in a model with induced inflammation, 451 proteins were detected in control in vitro culture, and 491 proteins were distinguished in samples of vestibular laminas of maxillary bone tissues originating from six different patients. Subsequent partial least squares discrimination analysis of obtained liquid chromatography-tandem mass spectrometry data was able to discriminate among in vitro cultures with induced cancer, with induced inflammation, and control cultivation. Thus, the direct in-sample protein digestion by trypsin followed by liquid chromatography-tandem mass spectrometry analysis of released specific peptide fragments from the insoluble matrix and mathematical analysis of the mass spectrometry data seems to be a promising tool for the routine proteomic characterization of in vitro human bone models with induced different pathological states. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2023 Electronic address https://doi.org/10.1002/jssc.202200694
Number of the records: 1