title: Humanization of yeast to produce complex terminally sialylated glycoproteins.
abstract: Yeast is a widely used recombinant protein expression system. We expanded its utility by engineering the yeast Pichia pastoris to secrete human glycoproteins with fully complex terminally sialylated N-glycans. After the knockout of four genes to eliminate yeast-specific glycosylation, we introduced 14 heterologous genes, allowing us to replicate the sequential steps of human glycosylation. The reported cell lines produce complex glycoproteins with greater than 90% terminal sialylation. Finally, to demonstrate the utility of these yeast strains, functional recombinant erythropoietin was produced.
authors.affiliation: GlycoFi Inc., 21 Lafayette Street, Suite 200, Lebanon, NH 03766, USA.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Hamilton
authors.forename: Robert C
authors.initials: RC
authors.lastname: Davidson
authors.forename: Natarajan
authors.initials: N
authors.lastname: Sethuraman
authors.forename: Juergen H
authors.initials: JH
authors.lastname: Nett
authors.forename: Youwei
authors.initials: Y
authors.lastname: Jiang
authors.forename: Sandra
authors.initials: S
authors.lastname: Rios
authors.forename: Piotr
authors.initials: P
authors.lastname: Bobrowicz
authors.forename: Terrance A
authors.initials: TA
authors.lastname: Stadheim
authors.forename: Huijuan
authors.initials: H
authors.lastname: Li
authors.forename: Byung-Kwon
authors.initials: BK
authors.lastname: Choi
authors.forename: Daniel
authors.initials: D
authors.lastname: Hopkins
authors.forename: Harry
authors.initials: H
authors.lastname: Wischnewski
authors.forename: Jessica
authors.initials: J
authors.lastname: Roser
authors.forename: Teresa
authors.initials: T
authors.lastname: Mitchell
authors.forename: Rendall R
authors.initials: RR
authors.lastname: Strawbridge
authors.forename: Jack
authors.initials: J
authors.lastname: Hoopes
authors.forename: Stefan
authors.initials: S
authors.lastname: Wildt
authors.forename: Tillman U
authors.initials: TU
authors.lastname: Gerngross
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000378:metabolism; D004921:Erythropoietin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006031:Glycosylation; D006801:Humans; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012794:Sialic Acids / Q000378:metabolism; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D011994:Recombinant Proteins; D012794:Sialic Acids; D012795:Sialoglycoproteins; D004921:Erythropoietin; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid