High performance anion exchange chromatographic and colorimetric methods for quality assessment of total and free polysaccharide content in type b conjugate vaccine containing lactose.

The presence of lactose as a stabilizer in type b (Hib) conjugate vaccine is a challenge for chromatographic resolution of its total and free poly ribosyl ribitol phosphate (PRP) content. Sample pretreatment using ultrafiltration was performed and had removed ≥95% of lactose in shorter time compared to the conventional dialysis process. Separation of free unconjugated PRP was performed using solid-phase extraction C cartridges.

Molecular size distribution assessment of Haemophilus influenzae vaccine containing lactose by HPAEC-PAD and colorimetric assays.

Molecular size distribution of Haemophilus influenzae type b (Hib) conjugate vaccine is an important indicator for its immunogenicity and stability. Molecular size distribution was evaluated by High-Performance Protein Chromatography on Sepharose CL-4B column, and fractions were pooled. The use of high flow rate, incorporation of a calibration standard with the injected buffer and pooling method yielded a superior assay compared to conventional pharmacopeial method.

Robust Assessment of Molecular Size Distribution of Meningococcal AC Vaccine.

The molecular size of meningococcal polysaccharides is an important physicochemical parameter that is related to immunogenicity and efficacy. A simple method for size-exclusion chromatography was developed, optimized, and applied for safe and rapid fractionation of meningococcal polysaccharide AC vaccine. Pooling of the fractions collected from size-exclusion chromatography was investigated and evaluated, rather than analyzing each fraction separately, for determining the percentages of meningococcal polysaccharide A and C that were eluted before the distribution coefficient of 0.