Remolding N-Glycan Structure in HEK293 Cells

Abstract:

Many of biopharmaceutical proteins are produced by mammalian cells because the proteins require correct folding and glycosylation. However，production of recombinant proteins in mammalian cells has some issues to be solved，including higher manufacturing cost and heterogeneity of glycans attached to the proteins. To overcome the issue，we here established a HEK293 cell lines disrupted Golgi alpha-1，2-mannosidase I. In double-KO cell line，which was knocked out both MAN1A1 and MAN1A2，the high mannose-type N-linked glycans were increased，whereas complex type glycans was greatly reduced. The glycans of an endogenous protein LAMP2 were sensitive to endoglycosidase-H treatment in double-KO cells，indicating that most of glycans on the proteins were high mannose-type glycans. Our results suggest that disruption of alpha-1，2-mannosidases in mammalian cells，especially double-KO cells，is useful for production of recombinant proteins with high mannose-type glycans.