A method for purification of bacterial R-type lipopolysaccharides (lipooligosaccharides).
A new gel filtration method was developed for purification of R-type lipopolysaccharides (lipooligosaccharides) from some nonenteric gram-negative bacteria, including Neisseria meningitidis, Haemophilus influenzae, and Bordetella pertussis. These wild-type lipooligosaccharides are poorly extractable by the phenol-chloroform-ether extraction method of C. Galanos, O. Luderitz, and O. Westphal [1969) Eur. J. Biochem. 9, 245-249) and therefore a new procedure was developed for their isolation. The lipooligosaccharides (LOS) were first extracted by hot phenol-water, treated with RNase, then disaggregated in deoxycholic acid, and purified by gel filtration on Sephadex G-75. By comparison the conventional hot phenol-water purification method using repeated ultracentrifugations yielded less LOS. The yield of LOS by gel filtration was 30 to 108% higher and the purity was better.
D001886:Bordetella pertussis / Q000032:analysis; D002850:Chromatography, Gel / Q000379:methods*; D006090:Gram-Negative Bacteria / Q000032:analysis*; D006193:Haemophilus influenzae / Q000032:analysis; D008070:Lipopolysaccharides / Q000302:isolation & purification*; D009345:Neisseria meningitidis / Q000032:analysis; D019800:Phenol; D010636:Phenols; D014867:Water
D008070:Lipopolysaccharides; D010636:Phenols; C023023:lipid-linked oligosaccharides; D014867:Water; D019800:Phenol