Chapter title |
Ultra-Sensitive, High-Resolution Liquid Chromatography Methods for the High-Throughput Quantitative Analysis of Bacterial Cell Wall Chemistry and Structure.
|
---|---|
Chapter number | 2 |
Book title |
Bacterial Cell Wall Homeostasis
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3676-2_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3674-8, 978-1-4939-3676-2
|
Authors |
Laura Alvarez, Sara B. Hernandez, Miguel A. de Pedro, Felipe Cava |
Editors |
Hee-Jeon Hong |
Abstract |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 41 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 9 | 22% |
Researcher | 8 | 20% |
Student > Doctoral Student | 4 | 10% |
Student > Bachelor | 2 | 5% |
Professor | 2 | 5% |
Other | 7 | 17% |
Unknown | 9 | 22% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 14 | 34% |
Agricultural and Biological Sciences | 10 | 24% |
Immunology and Microbiology | 4 | 10% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 2% |
Unspecified | 1 | 2% |
Other | 1 | 2% |
Unknown | 10 | 24% |