Summary
Lipopolysaccharide (LPS) is the signature glycolipid isolated from almost all Gram-negative bacteria. LPSs are well known for their ability to elicit the release of cytokines from eukaryotic cells including macrophages, neutrophils, and epithelial cells. LPS can be isolated free of contaminating nucleic acids and proteins by various techniques. In this review, we outline approaches for the isolation and preparation of LPSs for structural studies as well as preparation of very highly purified material for biological studies. Methods are also provided for the analysis of the purity and the structural composition of the LPSs. Finally, three methods for the isolation of lipid A are described.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Shear, M. J. (1941) Effect of concentrate from B. prodigiosus filtrate on subcutaneous primary induced mouse tumors.Cancer Res. 1, 732–741.
Luderitz, O., Staub, A. M., and Westphal, O. (1966) Immunochemistry of O and R antigens of Salmonella and related Enterobacteriaceae.Bacteriol. Rev. 30, 192–255.
Goldman, R. C. and Leive, L. (1980) Heterogeneity of antigenic-side-chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2. Eur. J. Biochem. 107, 145–153.
Palva, E. T. and Makela, P. H. (1980) Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Eur. J. Biochem. 107, 137–143.
Preston, A., Mandrell, R. E., Gibson, B. W., and Apicella, M. A. (1996) The lipooligosaccharides of pathogenic gram-negative bacteria. Crit. Rev. Microbiol. 22, 139–180.
Ribi, E., Haskins, W. T., Landy, M., and Milner, K. C. (1961) Preparation and host-reactive properties of endotoxin with low content of nitrogen and lipid. J. Exp. Med. 114, 647–663.
Galanos, C., Luderitz, O., and Westphal, O. (1969) A new method for the extraction of R lipopolysaccharides Eur. J. Biochem. 9, 245–249.
Roberts, N. A., Gray, G. W., and Wilkinson, S. G. (1967) Release of lipopolysaccharide during the preparation of cell walls of Pseudomonas aeruginosa. Biochim. Biophys. Acta 135, 1068–1071.
Leive, L. (1965) Release of lipopolysaccharide by EDTA treatment of E. coli. Biochem. Biophys. Res. Commun. 21, 290–296.
Goebel, W. F., Binkley, F., and Perlman, E. (1945) Studies on the Flexner group of dysentery bacilli. I. The specific antigens of Shigella paradysenteriae. J. Exp. Med. 81, 315–330.
Westphal, O. and Jann, K. (1965) Bacterial lipopolysaccharides. Extraction with phenol water and further applications of the procedure, in Methods in Carbohydrate Chemistry (Whistler, R. L., ed.), Academic Press, New York, pp. 83–91.
Darveau, R. P. and Hancock, R. E. W. (1983) Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J. Bacteriol. 155, 831–838.
Johnson, K. G., Perry, M. B., and McDonald, J. J. (1976) Studies of the cellular and free lipopolysaccharides from Neisseria canis and N. subflava. Can. J. Microbiol. 189, 189–196.
Hitchcock, P. J. and Brown, T. M. (1983) Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J. Bacteriol. 154, 269–277.
Westphal, O., Jann, K., and Himmelspach, K. (1983) Chemistry and immunochemistry of bacterial lipopolysaccharides as cell wall antigens and endotoxins. Prog. Allergy 33, 9–39.
Johnson, K. G. and Perry, M. B. (1976) Improved techniques for the preparation of bacterial lipopolysaccharides. Can. J. Microbiol. 22, 29–34.
Erwin, A. L., Munford, R. S., and Group TBPFS. (1989) Comparison of lipopolysacharides from Brazilian purpuric fever isolates and conjunctivitis isolates of Haemophilus influenzae biogroup aegyptius. J. Clin. Microbiol. 27, 762–767.
Inzana, T. J. (1983) Electrophoretic heterogeneity and int erstrain variation of the lipopolysaccharide of Haemophilus influenzae. J. Infect. Dis. 148, 492–499.
Hitchcock, P. J. (1984) Analyses of gonococcal lipopolysaccharide in whole-cell lysates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis: stable association of lipopolysaccharide with the major outer membrane protein (protein I) of Neisseria gonorrhoeae. Infect. Immun. 46, 202–212.
Inzana, T. J. and Pichichero, M. E. (1984) Lipopolysaccharide subtypes of Haemophilus influenzae type b from an outbreak of invasive disease. J. Clin. Microbiol. 20, 145–150.
Tsai, C.-M. and Frasch, C. E. (1982) A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal. Biochem. 119, 115–119.
Blake, M. S., Johnston, K. H., Russell-Jones, G. J., and Gotschlich, E. C. (1984) A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal. Biochem. 136, 175–179.
Hitchcock, P. J. and Brown, T. M. (1983) Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J. Bacteriol. 154, 269–277.
Kulshin, V. A., Zahringer, U., Lindner, B., et al. (1992) Structural characterization of the lipid A component of pathogenic Neisseria meningitidis. J. Bacteriol. 174, 1793–1800.
El Hamidi, A., Tirsoaga, A., Novikov, A., Hussein, A., and Caroff, M. (2005) Microextraction of bacterial lipid A: easy and rapid method for mass spectrometric characterization. J. Lipid Res. 46, 1773–1778.
Yi, E. C. and Hackett, M. (2000) Rapid isolation method for lipopolysaccharide and lipid A from Gram-negative bacteria. Analyst 125, 651–656.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Apicella, M.A. (2008). Isolation and Characterization of Lipopolysaccharides. In: DeLeo, F.R., Otto, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology™, vol 431. Humana Press. https://doi.org/10.1007/978-1-60327-032-8_1
Download citation
DOI: https://doi.org/10.1007/978-1-60327-032-8_1
Publisher Name: Humana Press
Print ISBN: 978-1-58829-740-2
Online ISBN: 978-1-60327-032-8
eBook Packages: Springer Protocols