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

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

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Laboratorio de Carbohidratos y Glicoconjugados, Departamento de Desarrollo Biotecnológico y División Producción, Facultades de Medicina/Química, Instituto de, Higiene, Av. Alfredo Navarro 3051, CP 11600, Montevideo, Uruguay.


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.

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