Antibody-based therapeutics with enhanced ADCC activity
Methods for producing antibody-based therapeutics with enhanced ADCC activity are disclosed. The enhanced ADCC activity is attributed to oligomannose-type N-glycans on the antibodies and Fc fusion proteins of the invention. Also disclosed are methods of using such antibody-based therapeutics for targeted killing of cells in a mammal, including therapeutic methods of treating cancers, autoimmune diseases and other diseases.
1. A method of producing a glycosylated monoclonal antibody or Fc fusion protein comprising:
(a) providing a monoclonal antibody or Fc fusion protein producing cell;
(b) culturing the cell in the presence of an amount of kifunensine sufficient to produce a monoclonal antibody or Fc fusion protein having at least 50% Man 5-9 (GlcNAc) 2 N-glycans, wherein Man8 and Man9-containing N-glycans together are the major species; and
(c) recovering the glycosylated antibody or Fc fusion protein.
2. The method of claim 1 , wherein the amount of kifunensine is less than 1000 ng/ml.
3. The method of claim 1 , wherein the amount of kifunensine is less than 500 ng/ml.
4. The method of claim 1 , wherein the amount of kifunensine is less than 100 ng/ml.
5. The method of claim 1 , wherein the amount of kifunensine is less than 80 ng/ml.
6. The method of claim 1 , wherein the amount of kifunensine is at least about 60 ng/ml.
7. The method of claim 1 , wherein the amount of kifunensine is about 60 to about 2500 ng/ml.
8. The method of claim 1 , wherein the monoclonal antibody or Fc fusion protein has at least 60% Man 5-9 (GlcNAc) 2 N-glycans.
9. The method of claim 1 , wherein the monoclonal antibody or Fc fusion protein has at least 70% Man 5-9 (GlcNAc) 2 N-glycans.
10. The method of claim 1 , wherein the monoclonal antibody or Fc fusion protein has at least 90% Man 5-9 (GlcNAc) 2 N-glycans.
11. The method of claim 1 , wherein the amount of kifunensine is sufficient to produce a monoclonal antibody or Fc fusion protein having less than 30% Man 5 (GlcNAc) 2 and/or Man 6 (GlcNAc) 2 N-glycans.
12. The method of claim 1 , wherein the amount of kifunensine sufficient to produce a monoclonal antibody or Fc fusion protein having less than 30% fucosylated N-glycans.