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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 universitet, Medicinska fakulteten, Molekylär Infektionsmedicin, Sverige (MIMS). Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten). Umeå universitet, Medicinska fakulteten, Umeå Centre for Microbial Research (UCMR).
Umeå universitet, Medicinska fakulteten, Molekylär Infektionsmedicin, Sverige (MIMS). Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten). Umeå universitet, Medicinska fakulteten, Umeå Centre for Microbial Research (UCMR).
Umeå universitet, Medicinska fakulteten, Molekylär Infektionsmedicin, Sverige (MIMS). Umeå universitet, Medicinska fakulteten, Institutionen för molekylärbiologi (Medicinska fakulteten). Umeå universitet, Medicinska fakulteten, Umeå Centre for Microbial Research (UCMR).
2016 (engelsk)Inngår i: Bacterial cell wall homeostasis: methods and protocols /edited by Hee-Jeon Hong / [ed] Hee-Jeon Hong, New York: Humana Press, 2016, Vol. 1440, s. 11-27Kapittel i bok, del av antologi (Fagfellevurdert)
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.

sted, utgiver, år, opplag, sider
New York: Humana Press, 2016. Vol. 1440, s. 11-27
Serie
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 1440
Emneord [en]
Cell wall, HPLC, Muropeptide, Peptidoglycan, Reverse-phase liquid chromatography, UPLC
HSV kategori
Identifikatorer
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 (tryckt)ISBN: 978-1-4939-3676-2 (digital)OAI: oai:DiVA.org:umu-128371DiVA, id: diva2:1051739
Tilgjengelig fra: 2016-12-02 Laget: 2016-12-02 Sist oppdatert: 2018-06-09bibliografisk kontrollert

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