Determination of Molecular Size Distribution of Meningococcal Polysaccharide Vaccine by Size Exclusion Chromatography Method

Abstract

Neisseria meningitidis, the leading cause of bacterial meningococcal disease has been considered as a serious public health threat worldwide due to its higher rate of morbidity and mortality. The capsular polysaccharides of these bacteria are potential vaccine candidates being efficacious to provide protection against meningococcal disease. Tetravalent vaccine is manufactured from distinct polysaccharides of serogroup A, C, W and Y that has been clinically proved to be a suitable vaccine against broad range of meningococcal infections. The molecular size distribution of polysaccharide vaccine is a critical parameter which is directly correlated with the immunogenicity. In the present study, the distribution coefficient value of meningococcal polysaccharide serogroup A, C, W and Y were evaluated by using a size exclusion chromatography method followed by a rapid and reliable quantitative rocket Immunoelectrophoresis assay. the analyses were performed by high performance liquid chromatography system AKTA Purifier 100 on a XK 16/70 (700mmx16mm) column packed with cross linked Sepharose CL-4B for determination of molecular size distribution of eight different batches of Ingovax® ACWY, a meningococcal polysaccharide vaccine produced by Incepta Vaccine Ltd, Bangladesh. The calculated mean distribution coefficient and standard deviation values of serogroups A, C, W and Y were found to be 0.28±0.14, 0.20±0.09, 0.19±0.11 and 0.14±0.12 respectively and met the requirements of British Pharmacopeia. The results also included the calculation and comparative analysis of distribution coefficient value of each serogroup. The method was proved to be robust for determining the distribution coefficient values which is an obligatory requirement for vaccine lot release. In fine, the study provided the information on distribution coefficient values of the meningococcal polysaccharide vaccine whether they comply with the recommendation of British Pharmacopeia.