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Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction
Published on: May 28, 2012
Purification of Lipopolysaccharides from Leptospira
Delphine Bonhomme1, Catherine Werts2
1Institut Pasteur, Biology and Genetic of the Bacterial Cell Wall Unit, Innate Immunity & Leptospira Group, Paris, France.
Abstract:
Leptospira species are one of the few spirochetes to possess a lipopolysaccharide (LPS) embedded in their outer membrane. Two protocols are currently available to extract and/or purify the leptospiral lipopolysaccharides: the rapid proteinase K method and the classical hot water/phenol extraction. The first method allows to get a quick overview of the LPS O antigen structure, whereas the second method is fitted to study the immunological properties of the leptospiral LPS. These two methods will be detailed in this chapter. Methodologies to assess the quality of the purification, such as the modified silver staining coloration, will also be reviewed. Both advantages and limitations of the different analyses will be described.

