Production of galactosylated glycoproteins in lower eukaryotes

摘要

The present invention provides a novel lower eukaryotic host cell producing human-like glycoproteins characterized as having a terminal β-galactose residue and essentially lacking fucose and sialic acid residues. The present invention also provides a method for catalyzing the transfer of a galactose residue from UDP-galactose onto an acceptor substrate in a recombinant lower eukaryotic host cell, which can be used as a therapeutic glycoprotein.

主权项

1. A method for producing a glycoprotein having one or more galactose residues in a Pichia pastoris host cell genetically engineered to produce N-linked oligosaccharides having terminal GlcNAc residues, the method comprising expressing in the host cell: a nucleic acid encoding the glycoprotein, a nucleic acid encoding a fusion protein comprising β-galactosyltransferase and a targeting peptide that targets the fusion protein to the endoplasmic reticulum (ER) or Golgi of the host cell, and a nucleic acid encoding a UDP-galactose C4 epimerase enzyme or a catalytically active fragment thereof, wherein the method results in the production of the glycoprotein having one or more galactose residues, and wherein at least 60 mole % of the N-glycans in the glycoprotein comprise the galactose residues.