Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1983 Apr;40(1):39–45. doi: 10.1128/iai.40.1.39-45.1983

Preparation and immunochemical characterization of meningococcal group C polysaccharide-tetanus toxoid conjugates as a new generation of vaccines.

E C Beuvery, F Miedema, R van Delft, J Haverkamp
PMCID: PMC264815  PMID: 6187693

Abstract

Neisseria meningitidis group C polysaccharide-tetanus toxoid conjugates have been prepared by using high-molecular-weight polysaccharide and purified tetanus toxoid and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide as a coupling reagent. The influence of three conditions of preparation was studied. Biochemical assays, the enzyme-linked immunosorbent assay, and isopycnic CsCl gradient ultracentrifugation have been used to characterize the conjugates. The polysaccharide-to-protein ratios of the various conjugate preparations showed differences. The ability of the group C polysaccharide component to react with specific antibodies was reduced, whereas most of the tetanus toxoid seemed to be hidden by the polysaccharide. The composition of the conjugate was not homogeneous, and at least 10% of free polysaccharide was present. Thermostability in lyophilized condition in the presence of lactose was excellent.

Full text

PDF
41

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson P., Insel R. A. A polysaccharide-protein complex from Haemophilus influenzae type b. I. Activity in weanling rabbits and human T lymphocytes. J Infect Dis. 1981 Dec;144(6):509–520. doi: 10.1093/infdis/144.6.509. [DOI] [PubMed] [Google Scholar]
  2. Anderson P., Insel R. A., Smith D. H., Cate T. R., Couch R. B., Glezen W. P. A polysaccharide-protein complex from Haemophilus influenzae type b. III. Vaccine trial in human adults. J Infect Dis. 1981 Dec;144(6):530–538. doi: 10.1093/infdis/144.6.530. [DOI] [PubMed] [Google Scholar]
  3. Beuvery E. C. Immunisation against bacterial meningitis. J Infect. 1981 Mar;3(1 Suppl):71–79. doi: 10.1016/s0163-4453(81)80011-4. [DOI] [PubMed] [Google Scholar]
  4. Beuvery E. C., Leussink A. B., Van Delft R. W., Tiesjema R. H., Nagel J. Immunoglobulin M and G antibody responses and persistence of these antibodies in adults after vaccination with a combined meningococcal group A and group C polysaccharide vaccine. Infect Immun. 1982 Aug;37(2):579–585. doi: 10.1128/iai.37.2.579-585.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beuvery E. C., van Rossum F., Nagel J. Comparison of the induction of immunoglobulin M and G antibodies in mice with purified pneumococcal type 3 and meningococcal group C polysaccharides and their protein conjugates. Infect Immun. 1982 Jul;37(1):15–22. doi: 10.1128/iai.37.1.15-22.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bhattacharjee A. K., Jennings H. J., Kenny C. P., Martin A., Smith I. C. Structural determination of the sialic acid polysaccharide antigens of Neisseria meningitidis serogroups B and C with carbon 13 nuclear magnetic resonance. J Biol Chem. 1975 Mar 10;250(5):1926–1932. [PubMed] [Google Scholar]
  7. Bizzini B., Blass J., Turpin A., Raynaud M. Chemical characterization of tetanus toxin and toxoid. Amino acid composition, number of SH and S-S groups and N-terminal amino acid. Eur J Biochem. 1970 Nov;17(1):100–105. doi: 10.1111/j.1432-1033.1970.tb01141.x. [DOI] [PubMed] [Google Scholar]
  8. Farquhar J. D., Hankins W. A., DeSanctis A. N., DeMeio J. L., Metzgar D. P. Clinical and serological evaluation of purified polysaccharide vaccines prepared from Neisseria meningitidis group Y. Proc Soc Exp Biol Med. 1977 Sep;155(4):453–455. doi: 10.3181/00379727-155-39828. [DOI] [PubMed] [Google Scholar]
  9. Farquhar J. D., Hankins W. A., Desanctis A. N., Demeio J. L., Metzgar D. P. Clinical and serological evaluation of a meningococcal polysaccharide vaccine groups A, C, and Y. Proc Soc Exp Biol Med. 1978 Jan;157(1):79–82. doi: 10.3181/00379727-157-39995. [DOI] [PubMed] [Google Scholar]
  10. Gotschlich E. C., Austrian R., Cvjetanović B., Robbins J. B. Prospects for the prevention of bacterial meningitis with polysaccharide vaccines. Bull World Health Organ. 1978;56(4):509–518. [PMC free article] [PubMed] [Google Scholar]
  11. Griffiss J. M., Brandt B. L., Altieri P. L., Pier G. B., Berman S. L. Safety and immunogenicity of group Y and group W135 meningococcal capsular polysaccharide vaccines in adults. Infect Immun. 1981 Dec;34(3):725–732. doi: 10.1128/iai.34.3.725-732.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hankins W. A., Gwaltney J. M., Jr, Hendley J. O., Farquhar J. D., Samuelson J. S. Clinical and serological evaluation of a meningococcal polysaccharide vaccine groups A, C, Y, and W135. Proc Soc Exp Biol Med. 1982 Jan;169(1):54–57. doi: 10.3181/00379727-169-41306. [DOI] [PubMed] [Google Scholar]
  13. Hoare D. G., Koshland D. E., Jr A method for the quantitative modification and estimation of carboxylic acid groups in proteins. J Biol Chem. 1967 May 25;242(10):2447–2453. [PubMed] [Google Scholar]
  14. Irwin G. R., Allen R. G., Segal H. G., Allen A. M., Putnak J. R., Cannon H. G., Top F. H., Jr Serodiagnosis of hepatitis B virus infection by antibody to core antigen. J Infect Dis. 1977 Jul;136(1):31–36. doi: 10.1093/infdis/136.1.31. [DOI] [PubMed] [Google Scholar]
  15. Jennings H. J., Lugowski C. Immunochemistry of groups A, B, and C meningococcal polysaccharide-tetanus toxoid conjugates. J Immunol. 1981 Sep;127(3):1011–1018. [PubMed] [Google Scholar]
  16. Kurzer F., Douraghi-Zadeh K. Advances in the chemistry of carbodiimides. Chem Rev. 1967 Apr;67(2):107–152. doi: 10.1021/cr60246a001. [DOI] [PubMed] [Google Scholar]
  17. Käyhty H., Karanko V., Peltola H., Sarna S., Mäkelä P. H. Serum antibodies to capsular polysaccharide vaccine of group A Neissera meningitidis followed for three years in infants and children. J Infect Dis. 1980 Dec;142(6):861–868. doi: 10.1093/infdis/142.6.861. [DOI] [PubMed] [Google Scholar]
  18. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  19. Lepow M. L., Goldschneider I., Gold R., Randolph M., Gotschlich E. C. Persistence of antibody following immunization of children with groups A and C meningococcal polysaccharide vaccines. Pediatrics. 1977 Nov;60(5):673–680. [PubMed] [Google Scholar]
  20. Lifely M. R., Gilbert A. S., Moreno C. Sialic acid polysaccharide antigens of Neisseria meningitidis and Escherichia coli: esterification between adjacent residues. Carbohydr Res. 1981 Aug 1;94(2):193–203. doi: 10.1016/s0008-6215(00)80717-x. [DOI] [PubMed] [Google Scholar]
  21. Nakane P. K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. doi: 10.1177/22.12.1084. [DOI] [PubMed] [Google Scholar]
  22. Peltola H., Mäkelä H., Käyhty H., Jousimies H., Herva E., Hällström K., Sivonen A., Renkonen O. V., Pettay O., Karanko V. Clinical efficacy of meningococcus group A capsular polysaccharide vaccine in children three months to five years of age. N Engl J Med. 1977 Sep 29;297(13):686–691. doi: 10.1056/NEJM197709292971302. [DOI] [PubMed] [Google Scholar]
  23. SVENNERHOLM L. Quantitative estimation of sialic acids. II. A colorimetric resorcinol-hydrochloric acid method. Biochim Biophys Acta. 1957 Jun;24(3):604–611. doi: 10.1016/0006-3002(57)90254-8. [DOI] [PubMed] [Google Scholar]
  24. Schneerson R., Barrera O., Sutton A., Robbins J. B. Preparation, characterization, and immunogenicity of Haemophilus influenzae type b polysaccharide-protein conjugates. J Exp Med. 1980 Aug 1;152(2):361–376. doi: 10.1084/jem.152.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tiesjema R. H., Beuvery E. C., te Pas B. J. Enhanced stability of meningococcal polysaccharide vaccines by using lactose as a menstruum for lyophilization. Bull World Health Organ. 1977;55(1):43–48. [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES