Polysaccharide purification for vaccine production using lytic enzymes, tangential flow filtration, and multimode chromatography
AssigneeInventprise, LLC
InventorsSubhash V. Kapre, Anup K. Datta
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AssigneeInventprise, LLC
InventorsSubhash V. Kapre, Anup K. Datta
Estimating the term…
An improved, cost effective and shortened process of purification of the capsular polysaccharide of S. pneumoniae, Group B Streptococcus, H. influenzae, S. typhoid and N. meningitidis is disclosed. The process includes a cocktail of enzyme treatment, tangential flow filtration, and multimodal chromatography purification. For Gram-negative bacteria, endotoxin removal process involves endotrap HD resin. This shortened process achieved the purity required by WHO/EP/BP for the use in human vaccine preparation, with simple steps and higher yield as compared to conventional processes. The steps of the process avoid use of organic solvents such as, for example, alcohol, phenol, and ultracentrifugation that are otherwise expensive and time consuming to perform, and/or health hazards for commercial use. This process disclosed is also simple, efficient, non-toxic, easy to scale-up, and environmentally friendly.
A polysaccharide composition obtained or derived from Gram-positive or Gram-negative bacteria, which has been treated with deoxycholate at a pH of about 3.5 to about 7.0, and an antimicrobial enzyme, wherein after being treated the composition contains 0.35 percent or less of nucleic acid and 0.85 percent or less of protein.
A polysaccharide composition obtained or derived from Gram-positive or Gram-negative bacteria, which has been treated with deoxycholate at a pH of about 3.5 to about 6.5, and an antimicrobial enzyme, wherein the antimicrobial enzyme comprises a sequential treatment of Benzonase, Mutanolysin/lysozyme, β-D-N-acetyl glucosaminidase and Proteinase K, and after being treated the composition contains 0.35 percent or less of nucleic acid and 0.85 percent or less of protein.
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