Bacterial Polysaccharide Purification via Alkaline Treatment
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Solution Overview
Problem
Current purification processes for Streptococcus pneumoniae serotype 3 polysaccharides are inefficient, laborious, and result in high impurity levels and reduced yield due to the viscosity of the polysaccharide chains and challenges in handling and separating contaminants like DNA, RNA, and proteins.
Innovation Solution
A method involving a base treatment step to disrupt protein-polysaccharide associations, followed by clarification through decantation, sedimentation, filtration, centrifugation, ultrafiltration, and activated carbon filtration, to achieve a purified polysaccharide solution with reduced impurities and improved yield.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional purification processes are used for Streptococcus pneumoniae serotype 3 polysaccharides, then the polysaccharide can be purified, but the process is laborious, inefficient, and results in reduced yield due to high viscosity and impurity levels
Solution Approach 1:
The patent changes the pH parameter by treating the polysaccharide solution with base (raising pH above 8.0, preferably above 10.0) to alter the physical and chemical properties of the polysaccharide, reducing viscosity and improving handling characteristics. This parameter change enables more efficient purification with reduced processing time and improved yield.
2Manufacturing precision
If base treatment is applied to disrupt protein-polysaccharide associations, then impurity removal is improved, but process complexity increases
Solution Approach 1:
The base treatment step changes the pH parameter to disrupt protein-polysaccharide associations, enabling better separation of impurities. This single parameter change (pH adjustment) achieves improved purification quality without requiring multiple complex process steps, as the alkaline conditions naturally facilitate impurity removal through subsequent clarification steps.
Solution Approach 2:
The patent replaces complex mechanical separation processes with chemical treatment (base addition) that naturally facilitates impurity removal. The chemical modification of the polysaccharide through base treatment creates conditions where impurities can be removed through simpler clarification steps rather than requiring complex mechanical separation systems.
3Manufacturing precision
If multiple clarification steps are used to remove impurities, then purity is improved, but processing time and complexity increase
Solution Approach 1:
The base treatment is applied as a preliminary action before clarification steps, modifying the polysaccharide and impurity mixture to facilitate subsequent separation. By raising the pH first, the polysaccharide properties are optimized for easier impurity removal, making the subsequent clarification steps more effective and potentially reducing the number of steps required while maintaining high purity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method provides a simple, scalable, and cost-effective purification process that significantly reduces impurities and increases the yield of purified Streptococcus pneumoniae serotype 3 polysaccharides, meeting quality standards and enhancing the production efficiency of vaccines.
Implementation Method 1
The solution can then be treated by a base to achieve a pH above 8.0, preferably a pH above 10.0. The base may be NaOH, KOH, LiOH, NaHCO3, Na2C03, KzC03, KCN, Et3N, NH3, HzN2H2, NaH, NaOMe, NaOEt or KOtBu.
Implementation Method 2
Following base treatment, the suspension can be clarified by decantation, sedimentation, filtration or centrifugation
Implementation Method 3
The Streptococcus pneumoniae serotype 3 polysaccharide containing solution can then be further clarified by Ultrafiltration and/or Diafiltration
Implementation Method 4
the solution may be further clarified by an activated carbon filtration step
Implementation Method 5
the Streptococcus pneumoniae serotype 3 polysaccharide containing solution may be further treated by a flocculation step
Data Source
AI summary
The present invention relates to methods for purifying bacterial polysaccharides, in particular for removing impurities from cellular lysates of bacteria producing polysaccharides.