Simple method for repurification of endotoxins for biological use.
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.
Tirsoaga
Alina
A
Equipe "Endotoxines," UMR 8619 du CNRS, IBBMC, Université de Paris-Sud, 91405 Orsay, France. Martine.Caroff@ibbmc.u-psud.fr
Novikov
Alexey
A
Adib-Conquy
Minou
M
Werts
Catherine
C
Fitting
Catherine
C
Cavaillon
Jean-Marc
JM
Caroff
Martine
M
D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007158:Immunologic Techniques; D008070:Lipopolysaccharides / Q000737:chemistry / Q000302:isolation & purification*; D016328:NF-kappa B / Q000032:analysis; D053474:Nod1 Signaling Adaptor Protein / Q000378:metabolism; D053473:Nod2 Signaling Adaptor Protein / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D051195:Toll-Like Receptor 2 / Q000502:physiology; D051197:Toll-Like Receptor 4 / Q000502:physiology
D004731:Endotoxins; D008070:Lipopolysaccharides; D016328:NF-kappa B; C119067:NOD1 protein, human; C429018:NOD2 protein, human; D053474:Nod1 Signaling Adaptor Protein; D053473:Nod2 Signaling Adaptor Protein; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4