Patent ID: 11965009
Assignee: TRUSTEES OF BOSTON UNIVERSITY
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A | IPC A  C

Claim 2:
3. A method for producing aggregate-free, full-length, monomeric diphtheria toxin fusion proteins comprising:
(a) transforming Corynebacterium diphtheria strain with a DNA vector comprising:
(i) a toxP;
(ii) a mutant toxO that blocks Fe-mediated regulation of gene expression;
(iii) a DNA sequence encoding a protein comprising:
a) a signal peptide;
b) diphtheria receptor binding domain or has a non-functional diphtheria toxin receptor binding domain; and
c) an IL-2 target receptor binding domain,
wherein the toxP and the mutant toxO regulate expression of the DNA sequence encoding the protein;

(b) incubating the transformed bacteria in a culture medium to allow expression of the protein and that is secreted into the culture medium; and
(c) purifying the diphtheria toxin fusion protein from the culture medium,
wherein the diphtheria toxin fusion protein is selected from the any one of SEQ ID NO:12, SEQ ID NO: 14 and SEQ ID NO:15.