Glycoconjugate Journal

, Volume 27, Issue 1, pp 69–77 | Cite as

Multimeric bivalent immunogens from recombinant tetanus toxin HC fragment, synthetic hexasaccharides, and a glycopeptide adjuvant

  • Aileen F. G. Bongat
  • Rina Saksena
  • Roberto Adamo
  • Yukari Fujimoto
  • Zenyu Shiokawa
  • Dwight C. Peterson
  • Koichi Fukase
  • Willie F. Vann
  • Pavol Kováč
Article

Abstract

Using recombinant tetanus toxin HC fragment (rTT-HC) as carrier, we prepared multimeric bivalent immunogens featuring the synthetic hexasaccharide fragment of O-PS of Vibrio cholerae O:1, serotype Ogawa, in combination with either the synthetic hexasaccharide fragment of O-PS of Vibrio cholerae O:1, serotype Inaba, or a synthetic disaccharide tetrapeptide peptidoglycan fragment as adjuvant. The conjugation reaction was effected by squaric acid chemistry and monitored in virtually real time by SELDI-TOF MS. In this way, we could prepare well-defined immunogens with predictable carbohydrate–carrier ratio, whose molecular mass and the amount of each saccharide attached could be independently determined. The ability to prepare such neoglycoconjugates opens unprecedented possibilities for preparation of conjugate vaccines for bacterial diseases from synthetic carbohydrates.

Keywords

Conjugate vaccine Vibrio cholerae Adjuvant Squaric acid Tetanus toxin C fragment 

Abbreviations

TT

Tetanus toxoid

rTT-HC

Recombinant tetanus toxin fragment C

HC

Tetanus toxin fragment C

O-PS

O-specific Polysaccharides

SELDI-TOF-MS

Surface-enhanced laser desorption time-of-flight mass spectrometry

BSA

Bovine serum albumin

NIS

N-Iodosuccinimide

PG

Peptidoglycan

Notes

Acknowledgement

This research was supported by the Intramural Research Program of the NIH, NIDDK.

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Aileen F. G. Bongat
    • 1
  • Rina Saksena
    • 3
  • Roberto Adamo
    • 1
  • Yukari Fujimoto
    • 2
  • Zenyu Shiokawa
    • 2
  • Dwight C. Peterson
    • 3
  • Koichi Fukase
    • 2
  • Willie F. Vann
    • 3
  • Pavol Kováč
    • 1
  1. 1.NIDDK, LBCNational Institutes of HealthBethesdaUSA
  2. 2.Department of Chemistry, Graduate School of ScienceOsaka UniversityToyonakaJapan
  3. 3.OVRR, CBER, FDALaboratory of Bacterial ToxinsBethesdaUSA

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