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Volume 55, Issue 10

Synthesis and characterization of alginate–tetanus toxoid conjugate Free

Abstract

Chronic infection with is the main proven perpetrator of lung function decline and ultimate mortality in cystic fibrosis (CF) patients. Mucoid strains of this bacterium elaborate mucoid exopolysaccharide, also referred to as alginate. Alginate-based immunization of naïve animals elicits opsonic antibodies and leads to clearance of mucoid from the lungs. Alginate was isolated from mucoid strain 8821M by repeated ethanol precipitation, dialysis, proteinase and nuclease digestion, and chromatography. To improve immunogenicity, the purified antigen was coupled to tetanus toxoid (TT) with adipic acid dihydrazide (ADH) as a spacer and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDAC) as a linker. The reaction mixture was passed through a Sepharose CL-4B column. The resulting conjugate was composed of TT and large-size alginate polymer at a ratio of about 3 : 1; it was non-toxic and non-pyrogenic, and elicited high titres of alginate-specific IgG. Antisera raised against the conjugate had high opsonic activity against the vaccine strain. The alginate conjugate was also able to protect mice against a lethal dose of mucoid . These data indicate that an alginate-based vaccine has significant potential to protect against chronic infection with mucoid in the CF host.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): ADH, adipic acid dihydrazide , CF, cystic fibrosis , EDAC, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide , i.p., intraperitoneal(ly) , KLH, keyhole-limpet haemocyanin , MEP, mucoid exopolysaccharide and TT, tetanus toxoid
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2006-10-01
2024-04-27
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Synthesis and characterization of Pseudomonas aeruginosa alginate–tetanus toxoid conjugate
J. Med. Microbiol. 55, 1441 (2006); https://doi.org/10.1099/jmm.0.46696-0
/content/journal/jmm/10.1099/jmm.0.46696-0
/content/journal/jmm/10.1099/jmm.0.46696-0
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