1887

Abstract

A rapid and efficient method for the preparation of highly pure meningococcal lipo-oligosaccharide (LOS) was developed. This used a Superose 6 column on a FPLC system to purify LOS from phenol-water extracts of cell lysates of The purest LOS preparations, with no detectable protein contamination and less than 0.5% (w/w) residual RNA, were obtained when cell lysates had been treated with RNase ONE before phenol extraction and chromatographic separation. Preparations that had received no ribonuclease treatment had 2-3% residual RNA contamination and predigestion of samples with RNase A, which only partially degraded the RNA present in the crude extracts, resulted in LOS samples contaminated with 15-20% residual RNA. The LOS purified from RNase ONE-treated extracts was highly endotoxic, and showed no reduction in antibody binding or specific endotoxin activity compared to unpurified material. Approximately 80% of the LOS applied to the chromatography column was recovered as purified material.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-1-57
1996-01-01
2024-12-06
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/1/mic-142-1-57.html?itemId=/content/journal/micro/10.1099/13500872-142-1-57&mimeType=html&fmt=ahah

References

  1. Borrebaeck C. A. K., Malmborg A.-C., Furebring C, Michaelsson A, Ward S, Danielsson L., Ohlin M. 1992; Kinetic analysis of recombinant antibody-antigen interactions: relation between structural domains and antigen binding. Biotechnology 10:697–698
    [Google Scholar]
  2. Brade H., Galanos C. 1982; Isolation, purification and chemical analysis of the lipopolysaccharide and lipid A of Acinetobacter calcoaceticus NCTC 10305. Eur J Biochem 122:233–237
    [Google Scholar]
  3. Brezincshek H. P., Brezincshek R. I., Lipsky P. E. 1995; Analysis of the heavy chain repertoire of human peripheral B cells using single-cell polymerase chain reaction. J Immunol 155:190–202
    [Google Scholar]
  4. Campagnari A. A., Spinola S. M., Lesse A. J., Abu Kwaik Y., Mandrell R. E., Apicella M. A. 1990; Lipooligosaccharide epitopes shared among Gram-negative non-enteric mucosal pathogens. Microb Pathog 8:353–362
    [Google Scholar]
  5. Darveau R. P., 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
    [Google Scholar]
  6. Dunn K., Virji M., Moxon E. R. 1995; Investigations into the molecular basis of meningococcal toxicity for human endothelial and epithelial cells: the synergistic effect of LPS and pili. Microb Pathog 18:81–96
    [Google Scholar]
  7. Eichberg J., Mokrasch L. C. 1969; Interference by oxidised lipids in the determination of protein by the Lowry procedure. Anal Biochem 30:386–390
    [Google Scholar]
  8. Estabrook M., Mandrell R. E., Apicella M. A., Griffiss J. M. 1990; Measurement of the human response to meningococcal lipooligosaccharide antigens by using serum to inhibit monoclonal antibody binding to purified lipooligosaccharide. Infect Immun 58:2204–2213
    [Google Scholar]
  9. Gamian A, Beurret M, Miction F., Brisson J. R., Jennings H. J. 1992; Structure of the L2 lipopolysaccharide core oligosaccharides of Neisseria meningitidis. J Biol Chem 267:922–925
    [Google Scholar]
  10. Griffiss J. M., Schneider H., Mandrell R. E., Yamasaki R., Jarvis G. A., Kim J. J., Gibson B. W., Hamadeh R., Apicella M. A. 1988; Lipooligosaccharides: the principal glycolipids of the neisserial outer membrane. Rev Infect Dis 10:S287–S295
    [Google Scholar]
  11. Gu X. X., Tsai C. M. 1991; Purification of rough-type lipopolysaccharides of Neisseria meningitidis from cells and outer membrane vesicles in spent media. Anal Biochem 196:311–318
    [Google Scholar]
  12. Gu X. X., Tsai C. M. 1993; Preparation, characterisation and immunogenicity of meningococcal lipooligosaccharide-derived oligosaccharide-protein conjugates. Infect Immun 61:1873–1880
    [Google Scholar]
  13. Gu X. X., Tsai C. M., Karpas A. B. 1992; Production and characterization of monoclonal antibodies to type 8 lipooligosaccharide of Neisseria meningitidis. J Clin Microbiol 30:2047–2053
    [Google Scholar]
  14. Herd J. K. 1971; Interference of hexosamines in the Lowry reaction. Anal Biochem 44:404–410
    [Google Scholar]
  15. Hitchcock P. J., Leive L, Makela H., Reitschel E. T., Strittmatter W., Morrison D. C. 1986; Lipopolysaccharide nomenclature past, present and future. J Bacteriol 166:699–705
    [Google Scholar]
  16. Inzana T. J. 1983; Electrophoretic heterogeneity and interstrain variation of the lipopolysaccharide of Haemophilus influenzae. J Infect Dis 148:492–499
    [Google Scholar]
  17. Inzana T. J., Seifert W. E., Williams R. P. 1985; Composition and antigenic activity of the oligosaccharide moiety of Haemophilus influenzae type b lipooligosaccharide. Infect Immun 48:324–330
    [Google Scholar]
  18. Janda J., Work E. 1971; A colorimetric estimation of lipopolysaccharides. FEBS Lett 16:343–345
    [Google Scholar]
  19. Jennings H. J., Bhattacharjee A. K., Kenne L, Kenny C. P., Calver G. 1980; The R-type lipopolysaccharides of Neisseria meningitidis. Can J Biochem 58:128–136
    [Google Scholar]
  20. Jennings H. J., Lugowski C., Ashton F. E. 1984; Conjugation of meningococcal lipopolysaccharide R-type oligosaccharides to tetanus toxoid as route to a potential vaccine against group B N. meningitidis. Infect Immun 43:407–412
    [Google Scholar]
  21. Johnson K. G., Perry M. B. 1976; Improved techniques for the preparation of bacterial lipopolysaccharides. Can J Microbiol 22:29–34
    [Google Scholar]
  22. Kalintsky G., Hummel J. P., Dierks C. 1959; Some factors which affect the enzymatic digestion of ribonucleic acid. J Biol Chem 234:1512–1516
    [Google Scholar]
  23. Kulshin V. A., Zahringer U, Lindner B, Frasch C. E., Tsai C. M., Dmitriev B. A., Rietschel E. T. 1992; Structural characterization of the lipid A component of pathogenic Neisseria meningitidis. J Bacteriol 174:1793–1800
    [Google Scholar]
  24. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  25. Malmborg A.-C., Michaelsson A, Ohlin M, Jansson B., Borrebaeck C. A. K. 1992; Real time analysis of antibody-antigen reaction kinetics. Scand J Immunol 35:643–650
    [Google Scholar]
  26. Mandrell R. E., Griffiss J. M., Macher B. A. 1988; Lipooligo-saccharides (LOS) of Neisseria gonorrhoeae and Neisseria meningitidis have components that are immunochemical similar to precursors of human blood group antigens. J Exp Med 168:107–126
    [Google Scholar]
  27. Markwell M. A., Haas S. M., Tolbert N. E., Bieber L. L. 1981; Protein determination in membrane and lipoprotein samples: manu l and automated procedures. Methods Enymol 12:296–303
    [Google Scholar]
  28. Meador J. I., Cannon B., Cannistraro V. J., Kennell D. 1990; Purification and characterisation of Escherichia coli RNase I: comparisons with RNase M. Eur J Biochem 187:549–553
    [Google Scholar]
  29. Morrison D. C. 1982; Bacterial endotoxins and pathogenesis. Rev Infect Dis 5:S733–S747
    [Google Scholar]
  30. Morrissey J. H. 1981; Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310
    [Google Scholar]
  31. Pavliak V., Brisson J.-R., Michon F., Uhrin D., Jennings H. J. 1993; Structure of the sialylated L3 lipopolysaccharide of Neisseria meningitidis. J Biol Chem 268:14146–14152
    [Google Scholar]
  32. Peltola H. 1983; Meningococcal disease: still with us. Rev Infect Dis 5:71–91
    [Google Scholar]
  33. Peppier M. S. 1984; Two physically and serologically distinct lipopolysaccharide profiles in strains of Bordetella pertussis and their phenotype variants. Infect Immun 43:224–232
    [Google Scholar]
  34. Perry M. B., Daoust V., Diena B. B., Ashton F. E., Wallace R. 1975; The lipopolysaccharides of Neisseria gonorrhoeae colony types 1 and 4. Can J Biochem 53:623–629
    [Google Scholar]
  35. Phillips N. J., John C. M., Reinders L. G., Gibson B. W., Apicella M. A., Griffiss J. M. 1990; Structural models for the cell surface lipooligosaccharides of Neisseria gonorrhoeae and Haemophilus influenzae. Biomed Environ Mass Spectrom 19:731–745
    [Google Scholar]
  36. Ray A, Readhead K, Selkirk S., Pool S. 1991; Variability in LPS composition, antigenicity and reactogenicity of phase variants of Bordetella pertussis. FEMS Microbiol Lett 79:211–218
    [Google Scholar]
  37. Rodahl E., Maeland J. A. 1984; Affinity chromatography for purification of antibodies to Neisseria gonorrhoeae and Neisseria meningitidis lipopolysaccharides. Acta Pathol Microbiol Immunol Scand Sect C 92:247–254
    [Google Scholar]
  38. Romanowska E. 1970; Sepharose gel filtration method of purification of lipopolysaccharides. Anal Biochem 33:383–389
    [Google Scholar]
  39. Romanowska E., Mulczyk M. 1968; Chemical studies on the specific fragment of Shigella sonnei phase II. Eur J Biochem 5:109–113
    [Google Scholar]
  40. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  41. Schneider H., Hale T. L., Zollinger W. D., Seid R. C., Hammack C. A., Griffiss J. M. 1984; Heterogeneity of molecular size and antigenic expression within lipooligosaccharides of individual strains of Neisseria gonorrhoeae and Neisseria meningitidis. Infect Immun 45:544–549
    [Google Scholar]
  42. Scholten R. J., Kuipers B., Valkenburg H. A., Dankert J., Zollinger W. D., Poolman J. T. 1994; Lipo-oligosaccharide immunotyping of Neisseria meningitidis by a whole-cell ELISA with monoclonal antibodies. J Med Microbiol 41:236–243
    [Google Scholar]
  43. Tsai C.-M., Frasch C. E. 1982; A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem 119:115–119
    [Google Scholar]
  44. Verheul A. F., Braat A. K., Leenhouts J. M., Hoogerhout P., Poolman J. T., Snippe H., Verhoef J. 1991; Preparation, characterization, and immunogenicity of meningococcal immuno-type L2 and L3,7,9 phosphoethanolamine group-containing oligosaccharide-protein conjugates. Infect Immun 59:843–851
    [Google Scholar]
  45. Westphal O., Jann K. 1965; Bacterial lipopolysaccharides: extraction with phenol-water and further applications of the procedure. Methods Carbohydr Chem 5:83–91
    [Google Scholar]
  46. Wu L., Tsai C.-M., Frasch C. E. 1987; A method for purification of bacterial R-type lipopolysaccharides (lipooligosaccharides). Anal Biochem 160:281–289
    [Google Scholar]
/content/journal/micro/10.1099/13500872-142-1-57
Loading
/content/journal/micro/10.1099/13500872-142-1-57
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error