Method for controlling
View Patent ↗The present invention provides improved methods for producing a solution containing high molecular weight isolated Streptococcus pneumoniae 19A capsular polysaccharides. In certain methods, a fermentation culture of Streptococcus pneumoniae bacterial cells that produce serotype 19A capsular polysaccharides is fermented for less than 6 hours before the bacterial cells are lysed the capsular polysaccharides are harvested. In other methods, CO 2 is supplied to the fermentation culture. Supplying CO 2 to the fermentation culture includes adding bicarbonate ions to the fermentation culture, adding carbonate ions to the fermentation culture, adding mixtures of bicarbonate and carbonate ions to the fermentation culture, and overlaying the fermentation culture with CO 2 .
1. A method for producing a solution containing high molecular weight isolated Streptococcus pneumoniae serotype 19A capsular polysaccharides, the method comprising:
a) preparing a fermentation culture of Streptococcus pneumoniae bacterial cells that produce serotype 19A capsular polysaccharides;
b) fermenting said fermentation culture for less than 6 hours;
c) lysing the bacterial cells in said fermentation culture; and
d) isolating Streptococcus pneumoniae serotype 19A capsular polysaccharides from said fermentation culture;
whereby a solution containing high molecular weight isolated Streptococcus pneumoniae serotype 19A capsular polysaccharides is produced.
2. The method of claim 1 , wherein said high molecular weight isolated Streptococcus pneumoniae serotype 19A capsular polysaccharide has a molecular weight of at least 480 kD.
3. The method of claim 1 , wherein step c) comprises fermenting said fermentation culture for less than 5 hours.
4. The method of claim 1 , wherein step c) comprises fermenting said fermentation culture for less than 4 hours.
5. The method of claim 1 , wherein step c) comprises fermenting said fermentation culture for between 3 hours and 6 hours.
6. A method for producing a solution containing high molecular weight isolated Streptococcus pneumoniae serotype 19A capsular polysaccharides, the method comprising:
a) preparing a fermentation culture of Streptococcus pneumoniae bacterial cells that produce serotype 19A capsular polysaccharides;
b) supplying CO 2 to said fermentation culture;
c) fermenting said fermentation culture for less than 6 hours;
d) lysing the bacterial cells in said fermentation culture; and
e) isolating Streptococcus pneumoniae serotype 19A capsular polysaccharides from said fermentation culture;
whereby a solution containing high molecular weight isolated Streptococcus pneumoniae serotype 19A capsular polysaccharides is produced.
7. The method of claim 6 , wherein said high molecular weight isolated Streptococcus pneumoniae serotype 19A capsular polysaccharide has a molecular weight of at least 480 kD.
8. The method of claim 6 , wherein step c) comprises fermenting said fermentation culture for less than 5 hours.
9. The method of claim 6 , wherein step c) comprises fermenting said fermentation culture for less than 4 hours.
10. The method of claim 6 , wherein step c) comprises fermenting said fermentation culture for between 3 hours and 6 hours.
11. The method of claim 6 , wherein supplying CO 2 to said fermentation culture comprises adding bicarbonate ion (HCO 3 − ) to the fermentation culture.
12. The method of claim 11 , wherein adding HCO 3 − to the fermentation culture comprises adding NaHCO 3 .
13. The method of claim 6 , wherein supplying CO 2 to said fermentation culture comprises adding carbonate ion (CO 3 2− ) to the fermentation culture.
14. The method of claim 13 , wherein adding CO 3 2− to the fermentation culture comprises adding Na 2 CO 3 .
15. The method of claim 6 , wherein supplying CO 2 to said fermentation culture comprises a first addition of NaHCO 3 and a second addition of Na 2 CO 3 .
16. The method of claim 6 , wherein supplying CO 2 to said fermentation culture comprises overlaying the fermentation culture with CO 2 .