Process for production diphtheria toxin
View Patent ↗The present invention provides a method of purifying diphtheria toxin comprising (1) fermenting a microorganism strain capable of producing diphtheria toxin using glucose as a carbon source, said method comprising adding glucose to a growing culture whereby the addition of glucose maintains microorganism growth effective to support diphtheria toxin production; and (2) purifying the diphtheria toxin from the culture by contacting a toxin containing preparation derived therefrom with an ion exchange matrix, eluting a fraction containing the toxin, applying the eluate to a hydrophobic matrix, and eluting a fraction containing the toxin.
1. A method for the production of diphtheria toxin wherein a microorganism capable of producing diphtheria toxin is fermented using glucose as a carbon source, said method comprising adding glucose to a growing culture whereby the addition of glucose maintains a microorganism growth effective to support diphtheria toxin production, wherein the fermentation medium comprises from about 0.5 to about 1.5% yeast extract.
2. The method as claimed in claim 1 wherein the diphtheria toxin is a mutated diphtheria toxin having a mutation in the B subunit.
3. The method as claimed in claim 2 wherein the mutated diphtheria toxin has a cytotoxic A subunit.
4. The method as claimed in claim 3 wherein the mutated diphtheria toxin is CRM 107.
5. The method as claimed in claim 1 wherein the microorganism is corynebacterium diphtheriae.
6. The method as claimed in claim 1 wherein glucose is added in a fed batch method.
7. The method as claimed in claim 6 wherein the pH is maintained at between 7.0 to about 7.5.
8. The method as claimed in claim 1 wherein the fermentation medium comprises no more than about 1% yeast extract.
9. The method as claimed in claim 1 wherein the fermentation medium comprises cystine.
10. The method as claimed in claim 1 , wherein the microorganism is E. coli.