Production of Therapeutic Proteins in Lower Eukaryotes

低等真核生物中治疗性蛋白质的生产

• 批准号: 6549759
• 负责人: STEFAN WILDT
• 金额: $ 10万
• 依托单位: GLYCOFI, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-15 至 2003-02-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6549759
• 关键词: Fungi; biotechnology; biotherapeutic agent; chimeric proteins; enzyme activity; genetic library; glycoproteins; glycosylation; high performance liquid chromatography; high throughput technology; interferon beta; mass spectrometry; polymerase chain reaction; protein engineering; recombinant proteins; technology /technique development

DESCRIPTION (provided by applicant): The pharmacological efficacy of a protein often depends on the protein's glycosylation structure, and successful manufacturing relies on host systems that produce glycoforms similar to those in humans. The overall objectives of this project are to develop tools for the metabolic engineering of fungal glycosylation pathways and to use these tools toward the creation of fungal strains capable of producing and secreting large quantities of recombinant therapeutic proteins with human-like glycosylation structures. The tool development strategy centers on a library-based approach that will allow rapid identification of properly targeted and expressed glycosylation enzymes in a high throughput manner. PROPOSED COMMERICIAL APPLICATIONS: The current method of glycoprotein production based on mammalian cell culture systems, specifically Chinese hamster ovary cell lines and mouse fibroblast/myeloma cell lines, presents manufacturers with the problems of low production levels, high cost, and safety. In its present state, protein production technology cannot keep up with research and development efforts, making manufacturing the growth-limiting factor for the biotechnology industry. With 84 biological drugs on the market and well over 500 in clinical development, the proposed project will generate technology toward the manufacturing of glycosylated proteins with high production levels in a safe and cost-effective manner, thereby alleviating the production bottleneck and allowing more products to reach the patients who need them.

描述（由申请人提供）：蛋白质的药理学功效通常取决于蛋白质的糖基化结构，并且成功的生产依赖于产生与人体中的糖型相似的糖型的宿主系统。该项目的总体目标是开发用于真菌糖基化途径的代谢工程的工具，并使用这些工具来创建能够产生和分泌大量具有类人糖基化结构的重组治疗蛋白的真菌菌株。该工具开发策略以基于文库的方法为中心，该方法将允许以高通量方式快速鉴定正确靶向和表达的糖基化酶。建议的商业应用：目前基于哺乳动物细胞培养系统，特别是中国仓鼠卵巢细胞系和小鼠成纤维细胞/骨髓瘤细胞系的糖蛋白生产方法，给生产商带来了生产水平低、成本高和安全性的问题。目前，蛋白质生产技术跟不上研究和开发工作的步伐，使制造业成为生物技术工业增长的限制因素。该项目目前有84种生物药物在市场上销售，超过500种处于临床开发阶段，该项目将以安全和具有成本效益的方式生产高产量的糖基化蛋白质，从而缓解生产瓶颈，使更多的产品能够到达需要它们的患者。

项目成果

A combinatorial genetic library approach to target heterologous glycosylation enzymes to the endoplasmic reticulum or the Golgi apparatus of Pichia pastoris.

一种组合遗传文库方法，将异源糖基化酶靶向巴斯德毕赤酵母的内质网或高尔基体。

• DOI: 10.1002/yea.1835
• 发表时间: 2011
• 期刊: Yeast (Chichester, England)
• 作者: Nett,JuergenH;Stadheim,TerranceA;Li,Huijuan;Bobrowicz,Piotr;Hamilton,StephenR;Davidson,RobertC;Choi,Byung-Kwon;Mitchell,Teresa;Bobrowicz,Beata;Rittenhour,Alissa;Wildt,Stefan;Gerngross,TillmanU
• 通讯作者: Gerngross,TillmanU