酵母人源化以生产复杂的末端唾液酸化糖蛋白。

Humanization of yeast to produce complex terminally sialylated glycoproteins.

作者信息

Hamilton Stephen R, Davidson Robert C, Sethuraman Natarajan, Nett Juergen H, Jiang Youwei, Rios Sandra, Bobrowicz Piotr, Stadheim Terrance A, Li Huijuan, Choi Byung-Kwon, Hopkins Daniel, Wischnewski Harry, Roser Jessica, Mitchell Teresa, Strawbridge Rendall R, Hoopes Jack, Wildt Stefan, Gerngross Tillman U

机构信息

GlycoFi Inc., 21 Lafayette Street, Suite 200, Lebanon, NH 03766, USA.

出版信息

Science. 2006 Sep 8;313(5792):1441-3. doi: 10.1126/science.1130256.

DOI:10.1126/science.1130256

PMID:16960007

Abstract

Yeast is a widely used recombinant protein expression system. We expanded its utility by engineering the yeast Pichia pastoris to secrete human glycoproteins with fully complex terminally sialylated N-glycans. After the knockout of four genes to eliminate yeast-specific glycosylation, we introduced 14 heterologous genes, allowing us to replicate the sequential steps of human glycosylation. The reported cell lines produce complex glycoproteins with greater than 90% terminal sialylation. Finally, to demonstrate the utility of these yeast strains, functional recombinant erythropoietin was produced.

摘要

酵母是一种广泛使用的重组蛋白表达系统。我们通过对巴斯德毕赤酵母进行工程改造来扩展其用途，使其能够分泌具有完全复杂的末端唾液酸化N-聚糖的人糖蛋白。在敲除四个基因以消除酵母特异性糖基化后，我们引入了14个异源基因，从而能够重现人糖基化的连续步骤。报道的细胞系产生的复合糖蛋白末端唾液酸化程度超过90%。最后，为了证明这些酵母菌株的实用性，生产了功能性重组促红细胞生成素。