Patent ID: 7863020

Claim:
A method for producing a recombinant sialylated glycoprotein in a Pichia pastoris host cell, the host cell selected or engineered to produce recombinant glycoproteins comprising a glycoform selected from the group consisting of Gal (1-4) GlcNAc (1-4) Man 3 GlcNAc 2 , the method comprising: (a) transforming into the host cell one or more nucleic acid molecules encoding a bifunctional UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase, an N-acetylneuraminate-9-phosphate synthase, and a CMP-Sialic acid synthase; (b) transforming into the host cell a nucleic acid molecule encoding a CMP-sialic acid transporter; and (c) transforming into the host cell a nucleic acid molecule encoding a 2,6-sialyltransferase catalytic domain fused to a cellular targeting signal peptide encoded by nucleotides 1-108 of the S. cerevisiae Mnn2 wherein nucleotides 1-108 encoding the S. cerevisiae Mnn2 have the nucleotide sequence shown by nucleotides 13 to 120 of SEQ ID NO:103; wherein, upon passage of the recombinant glycoprotein through the secretory pathway of the host cell, a recombinant sialylated glycoprotein comprising a glycoform selected from the group consisting of NANA (1-4) Gal (1-4) GlcNAc (1-4) Man 3 GlcNAc 2 glycoform is produced.