Abstract
Glycoconjugate-based vaccines have proved to be effective at producing long-lasting protection against numerous pathogens. Here, we describe the application of bacterial protein glycan coupling technology (PGCT) to generate a novel recombinant glycoconjugate vaccine. We demonstrate the conjugation of the Francisella tularensis O-antigen to the Pseudomonas aeruginosa carrier protein exotoxin A using the Campylobacter jejuni PglB oligosaccharyltransferase. The resultant recombinant F. tularensis glycoconjugate vaccine is expressed in Escherichia coli where yields of 3 mg l(-1) of culture were routinely produced in a single-step purification process. Vaccination of BALB/c mice with the purified glycoconjugate boosted IgG levels and significantly increased the time to death upon subsequent challenge with F. tularensis subsp. holarctica. PGCT allows different polysaccharide and protein combinations to be produced recombinantly and could be easily applicable for the production of diverse glycoconjugate vaccines.
Keywords:
Francisella tularensis; glycoconjugate; protein glycan coupling technology; vaccine.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ADP Ribose Transferases / immunology*
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ADP Ribose Transferases / metabolism
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Animals
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Antibodies, Bacterial / blood
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Bacterial Toxins / immunology*
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Bacterial Toxins / metabolism
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Bacterial Vaccines* / chemistry
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Bacterial Vaccines* / immunology
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Campylobacter jejuni / enzymology
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Escherichia coli / metabolism
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Exotoxins / immunology*
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Exotoxins / metabolism
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Female
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Francisella tularensis / immunology*
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Francisella tularensis / metabolism
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Glycosylation
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Hexosyltransferases / metabolism
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Immunoglobulin G / blood
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Membrane Proteins / metabolism
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Mice
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Mice, Inbred BALB C
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O Antigens / immunology*
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O Antigens / metabolism
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Pseudomonas aeruginosa / immunology
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Pseudomonas aeruginosa / metabolism
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Pseudomonas aeruginosa Exotoxin A
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Technology, Pharmaceutical
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Tularemia / immunology
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Tularemia / prevention & control*
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Vaccines, Conjugate* / chemistry
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Vaccines, Conjugate* / immunology
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Virulence Factors / immunology*
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Virulence Factors / metabolism
Substances
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Antibodies, Bacterial
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Bacterial Toxins
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Bacterial Vaccines
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Exotoxins
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Immunoglobulin G
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Membrane Proteins
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O Antigens
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Vaccines, Conjugate
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Virulence Factors
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Hexosyltransferases
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ADP Ribose Transferases
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dolichyl-diphosphooligosaccharide - protein glycotransferase