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

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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.

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Fig. 1
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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

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Acknowledgement

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

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Author notes

  1. Roberto Adamo

    Present address: , Novartis Vaccines and Diagnostics, s.r.l, Siena, Italy

Authors and Affiliations

  1. NIDDK, LBC, National Institutes of Health, Bethesda, MD, 20892-0815, USA

    Aileen F. G. Bongat, Roberto Adamo & Pavol Kováč

  2. Department of Chemistry, Graduate School of Science, Osaka University, Toyonaka, Osaka, 560-0043, Japan

    Yukari Fujimoto, Zenyu Shiokawa & Koichi Fukase

  3. OVRR, CBER, FDA, Laboratory of Bacterial Toxins, 8800 Rockville Pike, Bethesda, MD, 20892, USA

    Rina Saksena, Dwight C. Peterson & Willie F. Vann

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  1. Aileen F. G. Bongat
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  2. Rina Saksena
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Correspondence to Pavol Kováč.

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Bongat, A.F.G., Saksena, R., Adamo, R. et al. Multimeric bivalent immunogens from recombinant tetanus toxin HC fragment, synthetic hexasaccharides, and a glycopeptide adjuvant. Glycoconj J 27, 69–77 (2010). https://doi.org/10.1007/s10719-009-9259-4

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  • DOI: https://doi.org/10.1007/s10719-009-9259-4

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