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Applied and Environmental Microbiology, March 2007, p. 1803-1808, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02452-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Simple Method for Repurification of Endotoxins for Biological Use{triangledown}

Alina Tirsoaga,1,2 Alexey Novikov,1 Minou Adib-Conquy,3 Catherine Werts,3,{dagger} Catherine Fitting,3 Jean-Marc Cavaillon,3 and Martine Caroff1*

Equipe "Endotoxines," UMR 8619 du CNRS, IBBMC, Université de Paris-Sud, 91405 Orsay, France,1 Department of Physical Chemistry, University of Bucharest, 030018 Bucharest, Romania,2 Cytokines & Inflammation Unit, Institut Pasteur, 75015 Paris, France3

Received 19 October 2006/ Accepted 9 January 2007

A method for obtaining highly purified endotoxin (lipopolysaccharide [LPS]) in a few hours by repurification of commercial or laboratory preparations was devised. It avoids the use of phenol, which is not suitable for phenol-soluble lipopolysaccharides nor for some industrial purposes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization mass spectrometry analysis confirmed the integrity of the purified LPSs. The purified products did not activate Toll-like receptor 2 (TLR2), nuclear oligomerization domain 1 (NOD1), or NOD2 but did activate TLR4. Applied to different lipopolysaccharides, the method also improved their mass spectra, thus facilitating their structural analysis.

* Corresponding author. Mailing address: Equipe "Endotoxines," UMR 8619 du CNRS, IBBMC, Université de Paris-Sud, 91405 Orsay, France. Phone: 33 1 69 15 71 91. Fax: 33 1 69 85 37 15. E-mail: Martine.Caroff{at}ibbmc.u-psud.fr.

{triangledown} Published ahead of print on 19 January 2007.

{dagger} Present address: Unité de Bactériologie Moléculaire et Médicale, Institut Pasteur, 75015 Paris, France.

Applied and Environmental Microbiology, March 2007, p. 1803-1808, Vol. 73, No. 6
0099-2240/07/$08.00+0     doi:10.1128/AEM.02452-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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