Patent ID: 8999671
Filing Date: 2015-04-07
Classification: C07K,C12N,C12P,C12Y

Abstract:
1. A method for producing a recombinant sialylated glycoprotein in a Pichia pastoris host cell, the host cell selected or engineered to produce glycoproteins comprising a GalGlcNAcMan 5 GlcNAc 2 glycoform, the method comprising the step of transforming into the host cell: a. a nucleic acid encoding an enzyme having sialyltransferase activity, wherein the nucleic acid encodes a sialyltransferase enzyme comprising the amino acid sequence of SEQ ID NO:104 or a catalytic domain thereof; and b. a nucleic acid encoding a CMP-sialic acid transporter; and c. one or more nucleic acids encoding a CMP-sialic acid pathway consisting of: a bifunctional UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine Kinase, an N-acetylneuraminate-9-phosphate synthase, and a CMP-Sialic acid synthase; wherein upon passage of the recombinant glycoprotein through the Golgi apparatus of the host cell, a recombinant glycoprotein comprising a NANAGalGlcNAcMan