Microextraction of bacterial lipid A: easy and rapid method for mass spectrometric characterization |
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Authors: | El Hamidi Asmaa Tirsoaga Alina Novikov Alexey Hussein Ahmed Caroff Martine |
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Affiliation: | Equipe Endotoxines, Unité Mixte de Recherche 8619 du Centre National de la Recherche Scientifique, Institut de Biochimie, Biophysique Moléculaire et Cellulaire, Université de Paris-Sud, 91405 Orsay, France. |
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Abstract: | Endotoxins (lipopolysaccharides) are the main components of Gram-negative bacterial outer membranes. A quick and simple way to isolate their lipid region (lipid A) directly from whole bacterial cells was devised. This method using hot ammonium-isobutyrate solvent was applied to small quantities of cells and proved to be indispensable when a rapid characterization of lipid A structure by mass spectrometry was required. Biological activities of endotoxins are directly related to the lipid A structures, which vary greatly with cell growth conditions. This method is suitable for rough- and smooth-type bacteria and very efficient for screening variations in lipid A structures. Data are acquired in a few hours and avoid the use of phenol in extraction. |
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Keywords: | endotoxin lipopolysaccharide hydrolysis |
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