METHOD OF PRODUCING RECOMBINANT PROTEINS WITH MANNOSE-TERMINATED N-GLYCANS

摘要

We describe a method of expressing a recombinant protein comprising mannose-terminated N-glycans from a host cell, the method comprising: (a) introducing a nucleic acid encoding a recombinant protein into a Chinese Hamster Ovary (CHO) cell comprising a mutation in the GnT 1 gene (GenBank Accession Number AF343963) leading to loss of GnT 1 function; and (c) expressing the recombinant protein from the host cell, in which the expressed recombinant protein comprises a mannose-terminated glycan structure, and in which the method does not include a step of introducing functional GnT-I into the host cell. The method may be used for producing recombinant glucocerebrosidase with a mannose-terminated glycan structure, suitable for treatment or prevention of Gaucher's Disease.

主权项

1. A recombinant protein comprising mannose-terminated N-glycans expressed by a host cell line according to a method which includes steps of:
(a) introducing a nucleic acid encoding a recombinant protein into a host cell comprising a mutation in the GnT I gene leading to loss of GnT I function, wherein the host cell is a Chinese Hamster Ovary (CHO) cell selected with Ricinus communis agglutinin I (RCA-I) or a descendent thereof; and (b) expressing the recombinant protein from the host cell, wherein the expressed recombinant protein comprises mannose-terminated N-glycans and the method does not include a step of introducing functional GnT I into the host cell.