Evaluation of impact of temperature and pH alterations on the size and antigenicity of meningococcal serogroup A and X polysaccharides and conjugates.

The changes in the recommended storage conditions of the glycoconjugate vaccines against Neisseria meningitidis (Men) serogroup A and serogroup X can affect its activity or potency. Elevated temperature and the change in pH may result in the physical instability leading to the size degradation of the polysaccharide (PS) and subsequent loss of PS epitopes. Moreover, high temperature may also result in protein aggregation and altered tertiary structure of the protein in the conjugate. Consequently, the construction of a potent glycoconjugate is dependent on optimal temperature and pH. The changes in both these conditions can also affect the production of a capsular polysaccharide (PS) and its conjugation to a protein carrier and may also affect the integrity of the vaccine molecule including the maintenance of the protective epitopes. In our study we have used inhibition ELISA as a tool to assess the impact of temperature and pH alterations on the antigenicity of N. meningitidis serogroup A and X, PS and conjugates and their correlation with the size distribution analysis using high pressure size exclusion chromatography. The studies on pH alterations from 5 to 9 led to minimal impact on size and antigenicity of all antigens, however, an elevated temperature adversely impacted the antigen size as well as antigenicity to varying extent. Results indicate the higher stability of MenX PS and conjugate as compared to that for MenA counterparts at elevated temperatures. Furthermore, both the MenA and MenX conjugates appears to be more stable as compared to the corresponding PSs.