Patent ID: 7029872
Filing Date: 2006-04-18
Classification: A61P,C07K,C12N,C12P,C12Y

Abstract:
1. A method for producing a recombinant glycoprotein comprising an N-glycan structure that comprises a Man 5 GlcNAc 2 glycoform in a lower eukaryotic host cell that does not display alpha-1,6 mannosyltransferase activity with respect to the N-glycan on a glycoprotein, the method comprising the step of introducing into the host cell a nucleic acid encoding an alpha-1,2 mannosidase enzyme selected to have optimal activity in the ER or Golgi of said host cell, the enzyme comprising: (a) an alpha-1,2 mannosidase catalytic domain having optimal activity in said ER or Golgi at a pH between 5.1 and 8.0; fused to (b) a cellular targeting signal peptide not normally associated with the catalytic domain selected to target the mannosidase enzyme to the ER or Golgi apparatus of the host cell; whereby, upon passage of the recombinant glycoprotein through the ER or Golgi apparatus of the host cell, in excess of 30 mole % of the N-glycan structures attached thereto have a Man