A method for purification of bacterial R-type lipopolysaccharides (lipooligosaccharides) |
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| Authors: | Lihua Wu Chao-Ming Tsai Carl E. Frasch |
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| Affiliation: | 1. Food Science Department, FARAH, Faculty of Veterinary Medicine, University of Liège, Boulevard de Colonster 20, 4100 Liège, Belgium;2. Microbiology Unit, Catholic University of Louvain, Avenue Hippocrate B1.54.01, 1200 Brussels, Belgium;1. Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC), Universidad de Cantabria-CSIC-SODERCAN, Santander, España;2. Servicio de Microbiología, Hospital Universitario Marqués de Valdecilla y Fundación Instituto de Investigación Marqués de Valdecilla (IDIVAL), Santander, España;3. Departamento de Biología Molecular, Universidad de Cantabria, Santander, España |
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| Abstract: | 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. |
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