SpringerProtocols: Abstract: Purification of Capsular Polysaccharide

Purification of Capsular Polysaccharide

By: Qingling Yang³, Harold Jennings³

Abstract

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Meningococcus is an aerobic, fastidious, Gram-negative diplococcus that is found only in humans. Its cell wall has a cytoplasmic membrane, a peptidoglycan layer, and an outer membrane. Meningococci isolated from the bloodstream or the spinal fluid are almost always encapsulated. The capsules are polysaccharides, which consist of repeating sugar units (1,2). The capsular polysaccharide can be isolated, purified, and chemically and physically defined. Based on the chemical and immunological specificity of its capsular polysaccharides, Neisseria meningitidis has been classified serologically into groups A, B, C, 29E, H, I, K, L, W135, X, Y and Z (2). Of these, groups A, B, and C are responsible for approx 90% of meningococcal meningitis, and group B meningococcus is responsible for the majority of meningococcal disease in industrialized countries (1,2). The structures of the repeating units of the meningococcal polysaccharides have been elucidated and are shown in Table 1.

Table 1 Structures of the Capsular Polysaccharides of N. meningitidis

Group	Structure of repeating unit	Reference
A	→6)α-D-ManNAc(1-PO4→	*(8)*
	3
	↑
	OAc
B	→8)α-D-NeuNAc(2→	*(5)*
C	→9)α-D-NeuNAc(2→	*(5)*
	7/8
	↑
	OAc
H	→4)α-D-Gal (l→2) glycerol 3-PO₄→	*(6)*
L	→3)α-D-GlcNAc (l→3) β-D-GlcNAc (l→3) α-D-GlcNAc-1-PO₄→	*(7)*
X	→4)α-D-GlcNAc-1 -PO₄→	*(4)*
Y	→6)α-D-Glc (l→4) α-D-NeuNAc (2→ (OAc)	*(9)*
W135	→6)α-D-Gal (l→4) α-D-NeuNAc (2→	*(9)*
Z	→3)α-D-GalNAc (1→1) glycerol 3-PO₄→	*(10)*
29E	→7)β-D-KDO (l→3) α-D-GalNAc (l→	*(11)*

Affiliation(s): (3) Institute for Biological Science, National Research Council of Canada, Ottawa, ON, Canada

Book Title: Meningococcal Vaccines: Methods and Protocols

Series: Methods in Molecular Medicine | Volume: 66 | Pub. Date: Jun-25-2001 | Page Range: 41-47 | DOI: 10.1385/1-59259-148-5:41

Subject: Immunology

References

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1.	Gotschlich, E. C. (1984) Meningococcal meningitis, in Bacterial Vaccines (Germanier, E., ed.), Academic Press, New York, pp. 237–255.

2.	Jennings, H. J. (1990) Capsular polysaccharides as vaccine candidates. Curr. Top. Microbiol. Immunol. 150, 97–127.

3.	Gotschlich, E. C., Liu, T-Y, and Artenstein, M. S. (1969) Human immunity to the meningococcus. III. Preparation and immunochemical properties of the group A, group B and group C meningococcal polysaccharides. J. Exp. Med. 129, 1349–1365.

4.	Bundle, D. R., Jennings, H. J., and Kenny, C. P. (1974) Studies on the group-specific polysaccharide of Neisseria meningitidis serogroup X and an improved procedure for its isolation. J. Biol. Chem. 249, 4797–4801.

5.	Bhattacharjee, A. K., Jennings, H. J., Kenny, C. P., Martin, A., and Smith, I. C. P. (1975) Structural determination of the sialic acid polysaccharide antigens of Neisseria meningitidis serogroup B and serogroup C with carbon 13 nuclear magnetic resonance. J. Biol. Chem. 250, 1926–1932.

6.	Michon, F., Roy, R., Jennings, H. J., and Ashton, F. E. (1984) Structural elucidation of the capsular polysaccharide of Neisseria meningitidis group H. Can. J. Chem. 62, 1519–1524.

7.	Ashton, F. E., Ryan, A., Diena, B., and Jennings, H. J. (1983) A new serogroup (L) of Neisseria meningitidis. J. Clin. Microbiol. 17, 722–727.

8.	Bundle, D. R., Smith, I. C. P., and Jennings, H. J. (1974) Determination of the structure and conformation of bacterial polysaccharides by carbon-13 nuclear magnetic resonance. J. Biol. Chem. 249, 2275–2281.

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10.	Jennings, H. J., Rosell, K. G., and Kenny, C. P. (1979) Structural elucidation of the capsular polysaccharide antigen of Neisseria meningitidis serogroup Z using ¹³C nuclear magnetic resonance. Can. J. Chem. 57, 2902–2907.

11.	Bhattacharjee, A. K., Jennings, H. J., and Kenny, C. P. (1978) Strutural elucidation of the 3-deoxy-D-mannooctulosonic acid containing meningococcal 29e capsular polysaccharide antigen using carbon-13 nuclear magnetic resonance. Biochemistry. 17, 645–651.

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