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Merck
  • Reduction of free polysaccharide contamination in the production of a 15-valent pneumococcal conjugate vaccine.

Reduction of free polysaccharide contamination in the production of a 15-valent pneumococcal conjugate vaccine.

PloS one (2020-12-11)
Yoon Hee Whang, Soo Kyung Kim, Hyeseon Yoon, Seuk Keun Choi, Yeong Ok Baik, Chankyu Lee, Inhwan Lee
摘要

Glycoconjugate vaccines are vaccines in which a bacterial polysaccharide antigen is conjugated to a carrier protein to enhance immunogenicity by promoting T cell-dependent immune response. However, the free (unreacted) polysaccharides remaining after the conjugation process can inhibit the immunogenicity of a conjugate vaccine. Thus, we aimed to reduce the unbound free polysaccharides in the polysaccharide-protein conjugation process for the development of a new 15-valent pneumococcal conjugate vaccine (PCV15) by varying some factors that may affect the conjugation results such as polysaccharide/protein ratio, polysaccharide size, and concentration of a coupling agent in a conjugation reaction mixture. Concentrations of a coupling agent, carbodiimide (EDAC), and a carrier protein (CRM197) used in PCV15 production, during the conjugation process, had little effect on the content of free polysaccharides. However, the size of the polysaccharide was identified as the critical factor to control the free polysaccharide content, with an inverse relationship observed between the molecular weight of the polysaccharide and the residual free polysaccharide content after conjugation. Based on these results, a new PCV15 with low free polysaccharide contamination was produced and tested for immunogenicity using a rabbit model to show that it induces similar level of immune responses in rabbits compared to a comparator vaccine Prevnar13®.

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Sigma-Aldrich
1-氰基-4-二甲氨基吡啶四氟硼酸酯, organic cyanylating reagent
SAFC
CDAP
Sigma-Aldrich
21-Hydroxyprogesterone, ≥97% (HPLC)