Materials and methods for improved immunoglycoproteins

Abstract

Immunoglycoproteins, including antibodies, with improved ADCC and altered glycosylation patterns are provided. Also provided are cell culturing methods and media for producing such immunoglycoproteins, and therapeutic uses of such immunoglycoproteins.

1 - 11 . (canceled) 
     
     
         12 . A composition comprising recombinant immunoglycoproteins, wherein the recombinant immunoglycoprotein molecules each comprise an N-linked glycosylation site for linkage of oligosaccharide and a binding domain that specifically binds to CD37, wherein at least 60% of the N-linked oligosaccharides of the N-linked glycosylation site contain no fucose, and wherein said recombinant immunoglycoproteins have at least a 10-fold increase in antibody-dependent cellular cytotoxicity (ADCC) as compared to control immunoglycoproteins of the same encoded amino acid sequence produced in Chinese Hamster Ovary (CHO) cells. 
     
     
         13 . The composition of  claim 12 , wherein at least 80% of the N-linked oligosaccharides of the N-linked glycosylation site contain no fucose. 
     
     
         14 . The composition of  claim 12 , wherein at least 90% of the N-linked oligosaccharides of the N-linked glycosylation site contain no fucose. 
     
     
         15 . The composition of  claim 12 , wherein at least 40% of the N-linked oligosaccharides of the N-linked glycosylation site have a hexose content of ten hexoses. 
     
     
         16 . The composition of  claim 12 , wherein at least 40% of the N-linked oligosaccharides of the N-linked glycosylation site have a glucose content of one to three glucose molecules. 
     
     
         17 . The composition of  claim 12 , wherein said recombinant immunoglycoproteins have at least a 12-fold increase in antibody-dependent cellular cytotoxicity (ADCC) as compared to control immunoglycoproteins of the same encoded amino acid sequence produced in Chinese Hamster Ovary (CHO) cells. 
     
     
         18 . The composition of  claim 12 , wherein said recombinant immunoglycoproteins have at least an 18-fold increase in antibody-dependent cellular cytotoxicity (ADCC) as compared to control immunoglycoproteins of the same encoded amino acid sequence produced in Chinese Hamster Ovary (CHO) cells. 
     
     
         19 . The composition of  claim 12 , wherein said recombinant immunoglycoproteins are antibodies. 
     
     
         20 . The composition of  claim 12 , wherein said recombinant immunoglycoproteins each comprise a binding domain that specifically binds to CD37, an immunoglobulin hinge region peptide, and immunoglobulin CH2 and CH3 domains. 
     
     
         21 . The composition of  claim 12 , wherein said immunoglobulin hinge region peptide and immunoglobulin CH2 and CH3 domains are derived from a human IgG1. 
     
     
         22 . The composition of  claim 12 , wherein said recombinant immunoglycoproteins each comprise a binding domain that specifically binds to CD37 and immunoglobulin CH2 and CH3 domains. 
     
     
         23 . The composition of  claim 22 , wherein said N-linked glycosylation site is present in the CH2 domain. 
     
     
         24 . The composition of  claim 23 , wherein said N-linked glycosylation site is at Asn297 using Kabat numbering. 
     
     
         25 . The composition of  claim 12 , wherein said recombinant immunoglycoproteins have the same serum half-life as compared to control immunoglycoproteins of the same encoded amino acid sequence produced in Chinese Hamster Ovary (CHO) cells. 
     
     
         26 . The composition of  claim 12  formulated in a pharmaceutically acceptable carrier or diluent. 
     
     
         27 . A method of inhibiting cancer cell growth in a subject in need thereof comprising administering to the subject a composition of  claim 12 . 
     
     
         28 . The method of  claim 27 , wherein said composition is administered parenterally. 
     
     
         29 . The method of  claim 27 , wherein said composition is administered by direct injection into the tissue at the site of the cancer. 
     
     
         30 . The method of  claim 27 , wherein cancer cells expressing CD37 on their surface are depleted in the subject following administration of the composition.