This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kubler-Kielb, J.
Right arrow Articles by Schneerson, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kubler-Kielb, J.
Right arrow Articles by Schneerson, R.

 Previous Article  |  Next Article 

Infection and Immunity, August 2006, p. 4744-4749, Vol. 74, No. 8
0019-9567/06/$08.00+0     doi:10.1128/IAI.00315-06

Additional Conjugation Methods and Immunogenicity of Bacillus anthracis Poly-{gamma}-D-Glutamic Acid-Protein Conjugates

Joanna Kubler-Kielb,* Teh-Yung Liu, Christopher Mocca, Fathy Majadly, John B. Robbins, and Rachel Schneerson

Laboratory of Developmental and Molecular Immunity, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892

Received 24 February 2006/ Returned for modification 3 April 2006/ Accepted 28 April 2006

The capsule of Bacillus anthracis, composed of poly-{gamma}-D-glutamic acid ({gamma}DPGA), is an essential virulence factor of B. anthracis. The capsule inhibits innate host defense through its antiphagocytic action. {gamma}DPGA is a poor immunogen, but when covalently bound to a carrier protein, it elicits serum antibodies. To identify the optimal construct for clinical use, synthetic {gamma}DPGAs of different lengths were bound to carrier proteins at different densities. The advantages of the synthetic over the natural polypeptide are the homogeneous chain length and end groups, allowing conjugates to be accurately characterized and standardized and their chemical compositions to be related to their immunogenicities. In the present study, we evaluated, in addition to methods reported by us, hydrazone, oxime, and thioether linkages between {gamma}DPGA and several proteins, including bovine serum albumin, recombinant Pseudomonas aeruginosa exotoxin A, recombinant B. anthracis protective antigen (rPA), and tetanus toxoid (TT). The effects of the dosage and formulation on the immunogenicities of the conjugates were evaluated in mice. All conjugates were immunogenic. The optimal {gamma}DPGA chain length of 10 to 15 amino acids and the density, an average of 15 mol {gamma}DPGA per mol of protein, were confirmed. The thioether bond was the optimal linkage type, and TT and rPA were the best carriers. The optimal dosage was 1.2 to 2.5 µg of {gamma}DPGA per mouse, and adsorption of the conjugates onto aluminum hydroxide significantly increased the antibody response to the protein with a lesser effect on anti-{gamma}DPGA levels.

* Corresponding author. Mailing address: National Institutes of Health, NICHD/LDMI, 9000 Rockville Pike, Bldg. 6, Rm. 1A05, Bethesda, MD 20892. Phone: (301) 496-6141. Fax: (301) 480-3442. E-mail: kielbj{at}mail.nih.gov.

Editor: D. L. Burns

Infection and Immunity, August 2006, p. 4744-4749, Vol. 74, No. 8
0019-9567/06/$08.00+0     doi:10.1128/IAI.00315-06




This article has been cited by other articles:

  • Kubler-Kielb, J., Majadly, F., Wu, Y., Narum, D. L., Guo, C., Miller, L. H., Shiloach, J., Robbins, J. B., Schneerson, R. (2007). Long-lasting and transmission-blocking activity of antibodies to Plasmodium falciparum elicited in mice by protein conjugates of Pfs25. Proc. Natl. Acad. Sci. USA 104: 293-298 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»