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Ultra-sensitive, high-resolution liquid chromatography methods for the high-throughput quantitative analysis of bacterial cell wall chemistry and structure
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).
2016 (English)In: Bacterial cell wall homeostasis: methods and protocols /edited by Hee-Jeon Hong / [ed] Hee-Jeon Hong, New York: Humana Press, 2016, Vol. 1440, p. 11-27Chapter in book (Refereed)
Abstract [en]

High-performance liquid chromatography (HPLC) analysis has been critical for determining the structural and chemical complexity of the cell wall. However this method is very time consuming in terms of sample preparation and chromatographic separation. Here we describe (1) optimized methods for peptidoglycan isolation from both Gram-negative and Gram-positive bacteria that dramatically reduce the sample preparation time, and (2) the application of the fast and highly efficient ultra-performance liquid chromatography (UPLC) technology to muropeptide separation and quantification. The advances in both analytical instrumentation and stationary-phase chemistry have allowed for evolved protocols which cut run time from hours (2-3 h) to minutes (10-20 min), and sample demands by at least one order of magnitude. Furthermore, development of methods based on organic solvents permits in-line mass spectrometry (MS) of the UPLC-resolved muropeptides. Application of these technologies to high-throughput analysis will expedite the better understanding of the cell wall biology.

Place, publisher, year, edition, pages
New York: Humana Press, 2016. Vol. 1440, p. 11-27
Series
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 1440
Keywords [en]
Cell wall, HPLC, Muropeptide, Peptidoglycan, Reverse-phase liquid chromatography, UPLC
National Category
Microbiology Analytical Chemistry
Identifiers
URN: urn:nbn:se:umu:diva-128371DOI: 10.1007/978-1-4939-3676-2_2PubMedID: 27311661Scopus ID: 2-s2.0-84979988068ISBN: 978-1-4939-3674-8 (print)ISBN: 978-1-4939-3676-2 (electronic)OAI: oai:DiVA.org:umu-128371DiVA, id: diva2:1051739
Available from: 2016-12-02 Created: 2016-12-02 Last updated: 2018-06-09Bibliographically approved

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Alvarez, LauraHernandez, Sara BCava, Felipe
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Molecular Infection Medicine Sweden (MIMS)Department of Molecular Biology (Faculty of Medicine)Umeå Centre for Microbial Research (UCMR)
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