US 10,889,845 B2
Production of N-glycoproteins for enzyme assisted glycomodification
Shamim Herbert Rahman, Valby (DK); Carsten Behrens, Copenhagen N (DK); Malene Bech Vester-Christensen, Vedbæk (DK); Henrik Clausen, Holte (DK); Zhang Yang, Vanløse (DK); Adnan Fevzi Halim, Malmö (SE); and Eric Bennett, Kgs. Lyngby (DK)
Assigned to UNIVERSITY OF COPENHAGEN, Copenhagen (DK); and DANMARKS TEKNISKE UNIVERSITET, Kgs. Lyngby (DK)
Appl. No. 15/745,300
Filed by UNIVERSITY OF COPENHAGEN, Copenhagen K (DK); and DANMARKS TEKNISKE UNIVERSITET, Kgs. Lyngby (DK)
PCT Filed Jun. 15, 2016, PCT No. PCT/EP2016/063753
§ 371(c)(1), (2) Date Jan. 16, 2018,
PCT Pub. No. WO2017/008982, PCT Pub. Date Jan. 19, 2017.
Claims priority of provisional application 62/193,433, filed on Jul. 16, 2015.
Prior Publication US 2019/0203247 A1, Jul. 4, 2019
Int. Cl. A61K 48/00 (2006.01); C07H 21/04 (2006.01); C12P 21/00 (2006.01); C07K 14/505 (2006.01); C07K 14/61 (2006.01); C07K 16/00 (2006.01); C12N 15/90 (2006.01); C12N 9/64 (2006.01); C12N 5/00 (2006.01); C12N 5/071 (2010.01)
CPC C12P 21/005 (2013.01) [C07K 14/505 (2013.01); C07K 14/61 (2013.01); C07K 16/00 (2013.01); C12N 5/00 (2013.01); C12N 5/0682 (2013.01); C12N 9/6437 (2013.01); C12N 15/907 (2013.01); C07H 21/04 (2013.01); C07K 2317/41 (2013.01); C12N 2501/724 (2013.01); C12N 2510/02 (2013.01)] 5 Claims
 
1. A mammalian cell comprising one or more glycosyltransferase genes that have been partially or fully inactivated, and that has more homogeneous glycosylation capacities, wherein Alpha-1,6-Mannosyl-Glycoprotein 2-Beta-N-Acetylglucosaminyltransferase (mgat2) has been knocked out (KO), and, optionally wherein Alpha-1,3-Mannosyl-Glycoprotein 4-Beta-N-Acetylglucosaminyltransferase (mgat4A) and/or Alpha-1,3-Mannosyl-Glycoprotein 4-Beta-N-Acetylglucosaminyltransferase B (mgat4B) and/or Alpha-1,6-Mannosylglycoprotein 6-Beta-N-Acetylglucosaminyltransferase (mgat5) has been KO, allowing production of N-glycans with monoantennary structure, and further comprising KO of at least one sialyltransferase and KO of a UDP-G1cNAc:betaGa1 beta-1,3-N-acetylglucosaminyltransferase 2 (B3 gnt2) gene, allowing production of N-glycans with monoantennary structure and without sialic acid capping and without Poly-N-acetyllactosamine (poly-LacNAc).