Process for producing a capsular polysaccharide for use in conjugate vaccines
View Patent ↗A method for producing a polysaccharide and a conjugate vaccine including the polysaccharide produced according to the method. A characteristic step in the method is that the pH of the culture medium is kept at a constant value with base or acid until adjustment with respectively base or acid is not possible anymore. Using the method, capsular polysaccharide may be obtained in a high yield in a relatively short time. The method is straightforward, reproducible and cost-effective.
1. A method for recovering a polysaccharide from a fermentation broth, comprising:
employing a cationic detergent to precipitate the polysaccharide or part of the contaminants from the supernatant to obtain a first polysaccharide fraction;
employing alcohol to precipitate the polysaccharide from the first polysaccharide fraction to obtain a second polysaccharide fraction;
subjecting the second polysaccharide fraction to an alcohol precipitation in the presence of an anionic detergent, whereby the alcohol is present in a concentration which is below a concentration at which the polysaccharide precipitates;
precipitating the polysaccharide from the soluble fraction employing alcohol to obtain a polysaccharide precipitate; and
dissolving the polysaccharide precipitate and subjecting it to concentration and diafiltration.
2. The method according to claim 1 , wherein the anionic detergent comprises sodium deoxycholate.
3. The method according to claim 1 , wherein the anionic detergent has a final concentration of about 0.1-1% w/v.
4. The method according to claim 1 , wherein the alcohol comprises ethanol.
5. The method according to claim 1 , wherein the alcohol during the step of employing alcohol to precipitate the polysaccharide from the first polysaccharide fraction to obtain a second polysaccharide fraction has a final concentration of about 60-74% v/v.
6. The method according to claim 1 , wherein the alcohol during the step subjecting the second polysaccharide fraction to an alcohol precipitation in the presence of an anionic detergent has a final concentration of about 10-50% v/v.
7. The method according to claim 1 , wherein the alcohol employed during the step of precipitation of the polysaccharide from the soluble fraction has a final concentration of about 60-85% v/v.
8. The method according to claim 1 , wherein the cationic surfactant comprises hexadecyltrimethyl ammonium bromide.
9. The method according to claim 1 , wherein the polysaccharide is obtained from Haemophilus influenza type b.