N-Glycosylation

Abstract

The present invention relates to a mammalian cell comprising a gene encoding a polypeptide of interest, wherein the polypeptide of interest is expressed comprising one or more posttranslational modification patterns. These modifications are useful for example in glycoprotein production where the antibodies with the modifications have an enhanced antibody-dependent cell-mediated cytotoxicity (ADCC). The present invention also relates to methods for producing the glycoproteins and compositions comprising the glycoproteins, and their uses.

1 . (canceled) 
     
     
         2 . An isolated mammalian cell comprising homogeneous biantennary N-glycans, wherein mgat4B and mgat5 are inactivated in the cell, thereby generating the homogeneous biantennary N-glycans in the cell. 
     
     
         3 . The cell according to  claim 2 , wherein the genes are inactivated using zinc finger nucleases, TALENs, or CRISPR/Cas9. 
     
     
         4 . The cell according to  claim 2 , wherein Fut8 is additionally inactivated. 
     
     
         5 . The cell according to  claim 2 , wherein B3gnt2 is additionally inactivated. 
     
     
         6 . The cell according to  claim 2 , wherein Mgat2 is additionally inactivated. 
     
     
         7 . The cell according to  claim 2 , wherein st3gal3, st3gal4, and st3gal6 are additionally inactivated. 
     
     
         8 . The cell according to  claim 2 , wherein Mgat4a is additionally inactivated. 
     
     
         9 . The cell according to  claim 2 , wherein ST6GAL1 and/or ST6GAL2 is knocked in. 
     
     
         10 . The cell according to  claim 2 , wherein st3gal4 are additionally inactivated. 
     
     
         11 . The cell according to  claim 2 , wherein st3gal4, and st3gal6 are additionally inactivated. 
     
     
         12 . The cell according to  claim 2 , wherein B4galt1 is additionally inactivated. 
     
     
         13 . The cell according to  claim 12 , wherein Fut8 is additionally inactivated. 
     
     
         14 . The cell according to  claim 2 , wherein B4galt1 and B4galt3 are additionally inactivated. 
     
     
         15 . The cell according to  claim 14 , wherein Fut8 is additionally inactivated. 
     
     
         16 . The cell according to  claim 2 , wherein the cell is a CHO, NSO, SP2/0, YB2/0, CHO-K1, CHO-DXB11, CHO-DG44, CHO-S, HEK293, HUVEC, HKB, or PER-C6 cell. 
     
     
         17 . The cell according to  claim 16 , wherein the cell is a CHO cell. 
     
     
         18 . The cell according to  claim 2 , further comprising EPO, an IgG antibody, a protein involved in hemostasis, or a coagulation factor.