Engineering N-glycan Site Occupancy to Improve Recombinant Glycoprotein Production from Insect Cells

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The objective of this project is to enhance the co-translational attachment of N-linked oligosaccharides to polypeptides in insect cells with the goal of improving recombinant glycoprotein yields. One of the major limitations of the baculovirus-insect cell gene expression system is its inability to properly glycosylate heterologous glycoproteins. The investigators have hypothesized that during the process of N-linked oligosaccharide attachment in insect cells, a limitation may exist in the production of the lipid linked donor substrate, and/or the catalytic enzyme, oligosaccharide tranferase. The investigators will determine which of these two factors may be the limiting step. Recombinant DNA techniques will then be used to correct the limitation. The investigators will also examine whether reduced N-glycan cleavage may alleviate the problem of under-glycosylation.