US2021087587A1PendingUtilityA1

N-Glycosylation

Assignee: UNIV COPENHAGENPriority: Dec 12, 2014Filed: Nov 5, 2020Published: Mar 25, 2021

Est. expiryDec 12, 2034(~8.4 yrs left)· nominal 20-yr term from priority

Inventors:Henrik Clausen Zhang Yang Adnan Fevzi Halim Eric Paul Bennett Carsten Behrens Malene Bech Vester-Christensen Shamim Herbert Rahman

C12P 21/005C12N 2501/24C07K 14/505C07K 16/464C12N 2511/00C12N 2501/724C07K 2317/56C12N 15/907C07K 2317/24

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Claims

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.

Claims

exact text as granted — not AI-modified

1 . (canceled) 
     
     
         2 . A mammalian cell, wherein the sialyltransferase genes, St3gal4 and St3gal6 are inactivated 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 B3gnt2 is additionally inactivated. 
     
     
         5 . The cell according to  claim 2 , wherein Fut8 is additionally inactivated. 
     
     
         6 . The cell according to  claim 2 , wherein mgat4A/4B and/or mgat5 are additionally inactivated. 
     
     
         7 . The cell according to  claim 6 , wherein mgat2 is additionally knocked out. 
     
     
         8 . The cell according to  claim 6 , wherein ST6GAL1 and/or ST6GAL2 have been knocked in. 
     
     
         9 . The cell according to  claim 2 , wherein mgat4B and mgat5 are additionally inactivated 
     
     
         10 . The cell according to  claim 9 , wherein mgat2 is additionally knocked out. 
     
     
         11 . The cell according to  claim 9 , wherein ST6GAL1 and/or ST6GAL2 have been knocked in. 
     
     
         12 . The cell according to  claim 2 , wherein mgat2 is additionally inactivated. 
     
     
         13 . The cell according to  claim 12 , wherein ST6GAL1 and/or ST6GAL2 has been knocked in. 
     
     
         14 . The cell according to  claim 2  wherein ST6GAL1 and/or ST6GAL2 has been knocked in. 
     
     
         15 . The cell according to  claim 14 , wherein B3gnt2 is additionally inactivated. 
     
     
         16 . The cell according to  claim 2 , wherein mgat5 is additionally inactivated. 
     
     
         17 . The cell according to  claim 16 , wherein the ST6GAL1 and/or ST6GAL2 has been knocked in. 
     
     
         18 . The cell according to  claim 2 , wherein cosmc is additionally inactivated. 
     
     
         19 . The cell according to  claim 2 , wherein the cell is a CHO, NS0, SP2/0, YB2/0, CHO-K1, CHO-DXB11, CHO-DG44, CHO-S, HEK293, HUVEC, HKB, or PER-C6 cell. 
     
     
         20 . The cell according to  claim 9 , wherein the cell is a CHO cell. 
     
     
         21 . The cell according to  claim 2 , further comprising EPO, an IgG antibody, a protein involved in hemostasis, or a coagulation factor.

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