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

该项目的目的是增强昆虫细胞中N-连接低聚糖与多肽的共翻译附着，以提高重组糖蛋白的产量。 杆状病毒-昆虫细胞基因表达系统的主要限制之一是其不能适当地糖基化异源糖蛋白。 研究人员推测，在昆虫细胞中N-连接寡糖附着的过程中，脂质连接供体底物和/或催化酶寡糖转移酶的产生可能存在限制。 研究人员将确定这两个因素中的哪一个可能是限制步骤。 然后将使用重组DNA技术来纠正限制。 研究人员还将检查减少N-聚糖切割是否可以缓解糖基化不足的问题。

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.