Improved conjugation and purification strategies for the preparation of protein-polysaccharide conjugates.
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.
Suárez
N
N
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.
Massaldi
H
H
Franco Fraguas
L
L
Ferreira
F
F
D000818:Animals; D016667:Bacterial Capsules / Q000276:immunology* / Q000302:isolation & purification*; D001428:Bacterial Vaccines / Q000302:isolation & purification*; D005260:Female; D006001:Glycoconjugates / Q000138:chemical synthesis* / Q000302:isolation & purification*; D051379:Mice; D012710:Serum Albumin, Bovine; D013296:Streptococcus pneumoniae / Q000276:immunology*
D001428:Bacterial Vaccines; D006001:Glycoconjugates; D012710:Serum Albumin, Bovine