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: 6792130
• 负责人: STEFAN WILDT
• 金额: $ 23.83万
• 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 2005-02-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6792130
• 关键词: Drosophilidae; Golgi apparatus; Moniliales; alpha 1 antitrypsin; bioengineering /biomedical engineering; biotechnology; biotherapeutic agent; chimeric proteins; drug design /synthesis /production; endodeoxyribonuclease; endoplasmic reticulum; fermentation; glycoproteins; glycosylation; high performance liquid chromatography; high throughput technology; interferon beta; mannosidase; matrix assisted laser desorption ionization; peptide library; protein engineering; protein purification; recombinant proteins; technology /technique development

DESCRIPTION (provided by applicant): Through metabolic engineering of the yeast glycosylation pathway, GlycoFi is developing yeast capable of secreting recombinant proteins with human-like glycosylation. In Phase I, a toolkit of cloning vectors, strains, libraries and protocols were developed for targeting glycosylation enzymes to the yeast secretory pathway. The feasibility of this approach was demonstrated by the functional expression of sigma-1,2,-mannosidase in vivo, resulting in the production of a recombinant protein with 65 percent human-like Man5glycosylation structure. This Phase II proposal aims to develop an automated, high-throughput approach capable of screening 3000 samples per week for highly efficient enzyme catalytic domains. These enzymes will be incorporated into a production strain generating commercially important proteins with a human-like Man5glycosylation structure. Proposed Commercial Application: A shortage of manufacturing capacity has emerged as the single largest barrier preventing therapeutic proteins in the development pipeline from reaching the clinic. GlycoFi's novel yeast-based protein production platform will alleviate this bottleneck and allow the bio-pharmaceutical industry to bring their products to market in a faster and more cost effective manner.

描述（由申请人提供）：通过酵母糖基化途径的代谢工程，GlycoFi正在开发能够分泌具有类人糖基化的重组蛋白的酵母。在第一阶段，开发了一套克隆载体、菌株、文库和方案，用于将糖基化酶靶向酵母分泌途径。该方法的可行性通过σ-1，2-甘露糖苷酶在体内的功能性表达得到了证明，导致产生具有65%的类人Man 5糖基化结构的重组蛋白。该第二阶段提案旨在开发一种自动化的高通量方法，能够每周筛选3000个样品以获得高效的酶催化结构域。将这些酶掺入生产菌株中，产生具有类似人的Man 5糖基化结构的商业上重要的蛋白质。建议的商业应用：制造能力的短缺已经成为阻止开发管道中的治疗性蛋白质到达临床的最大障碍。GlycoFi的新型酵母蛋白生产平台将缓解这一瓶颈，并使生物制药行业能够以更快、更具成本效益的方式将其产品推向市场。