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

Claim 5:
6. A method for producing an aggregate-free, full-length, monomeric diphtheria toxin fusion proteins comprising:
(a) transforming Cornyebacterium diptheria strain with a DNA expression vector comprising:
(i) a toxP;
(ii) a mutant toxO that blocks Fe-mediated regulation of gene expression; and
(iii) a DNA sequence encoding a protein comprising:
a) signal peptide;
b) diptheria receptor binding domain or has a non-functional diptheria receptor binding domain, and
c) a target receptor binding domain selected from the group comprising IL-3, IL-4, IL-6, IL-7, IL-15, EGF, FGF, substance P, CD4, αMSH, GRP, TT fragment C, GCSF, heregulin β1, TNFα, TGFβ, a functional part thereof, or a combination thereof;
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 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.