	US 12,241,108 B2
	Genetically engineered eukaryotic cells producing LacNAc-glycoproteins
Nico Callewaert, Nevele (BE); Wander Van Breedam, Antwerp (BE); and Francis Santens, Brussels (BE)
Assigned to VIB VZW, Ghent (BE); and Universiteit Gent, Ghent (BE)
Appl. No. 16/646,889
Filed by VIB VZW, Ghent (BE); and UNIVERSITEIT GENT, Ghent (BE)
PCT Filed Sep. 13, 2018, PCT No. PCT/EP2018/074784 § 371(c)(1), (2) Date Mar. 12, 2020, PCT Pub. No. WO2019/053145, PCT Pub. Date Mar. 21, 2019.
Claims priority of application No. 1714764 (GB), filed on Sep. 14, 2017; and application No. 18171657 (EP), filed on May 9, 2018.
Prior Publication US 2020/0255878 A1, Aug. 13, 2020
Int. Cl. C12P 21/00 (2006.01); C12N 15/52 (2006.01)

CPC C12P 21/005 (2013.01) [C12N 15/52 (2013.01); C12Y 204/01038 (2013.01)]

3 Claims

1. A mammalian cell comprising:

a first exogenous nucleic acid sequence encoding an endoglucosaminidase enzyme wherein the endoglucosaminidase enzyme is operably linked to a secretion signal or the endoglucosaminidase is linked to an ER or Golgi localization signal, and

a second exogenous nucleic acid sequence encoding a glycoprotein,

wherein the mammalian cell is deficient in at least one of the following enzymes: UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE), N-acetylneuraminate-9-phosphate synthase, CMP-sialic acid synthase, CMP-sialic acid transporter, and sialyltransferase;

wherein the mammalian cell produces no detectable N-glycans comprising sialic acid residues; and

wherein the N-glycans on the glycoprotein consist of more than 90% LacNAc residues.