Improved conjugation and purification strategies for the preparation of protein-polysaccharide conjugates

J Chromatogr A. 2008 Dec 12;1213(2):169-75. doi: 10.1016/j.chroma.2008.10.030. Epub 2008 Oct 15.


A glycoconjugate constituted by the Streptococcus pneumoniae serotype 14 capsular polysaccharide (CPS14) and bovine serum albumin (BSA) was prepared, and the unique properties of Sephadex LH-20 were used to separate the conjugate from the unconjugated material. The strength of this approach consists in its capacity to produce pure polysaccharide-protein conjugate in good yield and free from unconjugated material, a common residual contaminant of this type of immunobiologicals. The CPS14-BSA conjugate prepared via an improved 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP)-activation technique was characterized chemically and its immunogenicity was evaluated in mice. The purified conjugate, unlike the corresponding polysaccharide, produced a T-cell-dependent response in this species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Capsules / immunology*
  • Bacterial Capsules / isolation & purification*
  • Bacterial Vaccines / isolation & purification*
  • Female
  • Glycoconjugates / chemical synthesis*
  • Glycoconjugates / isolation & purification*
  • Mice
  • Serum Albumin, Bovine
  • Streptococcus pneumoniae / immunology*


  • Bacterial Vaccines
  • Glycoconjugates
  • Serum Albumin, Bovine