High-performance reverse phase chromatography with fluorescence detection assay for characterization and quantification of pneumococcal polysaccharides.

Leonardo Canaán-Haden; José Cremata; Janoi Chang; Yury Valdés; Félix Cardoso; Vicente Vérez Bencomo

doi:10.1016/j.vaccine.2005.01.128

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High-performance reverse phase chromatography with fluorescence detection assay for characterization and quantification of pneumococcal polysaccharides.

Leonardo Canaán-Haden·José Cremata·Janoi Chang·Yury Valdés·Félix Cardoso·Vicente Vérez Bencomo

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Abstract

A methods using high-performance reverse phase (RP) chromatography with fluorescence detection, has been developed to determine the composition and identity of Streptococcus pneumoniae capsular polysaccharide used in formulating conjugate vaccine for prevention of pneumococcal infection. For the monosaccharide composition, the polysaccharides were subjected to hydrofluoric acid (HF) hydrolysis followed by trifluoroacetic acid (TFA). After acid hydrolysis, the released monosaccharides were re-N-acetylated and labeled with 2-aminobenzamide (2AB) by reductive amination reaction. High-performance RP chromatography was performed on C18 TSKODS 120T column. Nuclear magnetic resonance was used to confirm chemical structure and purity of pneumococcal capsular polysaccharides.

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Leonardo Canaán-Haden, José Cremata, Janoi Chang, Yury Valdés, Félix Cardoso, Vicente Vérez Bencomo. 2006-04-12. High-performance reverse phase chromatography with fluorescence detection assay for characterization and quantification of pneumococcal polysaccharides.. https://doi.org/10.1016/j.vaccine.2005.01.128

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