PRODUCTION OF ALPHA-GALACTOSIDASE A--TRANSFECTED PLANTS

α-半乳糖苷酶 A 的生产——转染植物

• 批准号: 2422447
• 负责人: THOMAS H. TURPEN
• 金额: $ 10万
• 依托单位: LARGE SCALE BIOLOGY CORPORATION
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-15 至 1998-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2422447
• 关键词: Fabry's disease; affinity chromatography; alpha galactosidase; biotechnology; drug design /synthesis /production; gene targeting; genetically modified plants; protein purification; protein sequence; recombinant proteins; transfection; transfection /expression vector

Successful development of affordable molecular therapies will require fundamentally new approaches to simultaneous production and bioprocessing of recombinant proteins. For example, metabolic storage disorders are the most common group of hereditary abnormalities and many of these conditions are likely to be treatable by supplementation with exogenously produced enzymes. Yet for any particular molecular defect, small numbers of individuals are affected and therefore projected treatment costs are very high. Fabry disease is a recessive X-linked deficiency in the catabolic activity of the lysosomal hydrolase alpha-galactosidase A (Ga1-A) and is the second most common hereditary metabolic storage disorder of man. The overall objective of this proposal is to determine the feasibility of economically producing custom-designed recombinant Ga1-A for therapeutic treatment of Fabry disease. Ga1-A from human sources has a homodimeric glycoprotein structure with a subunit molecular weight of approximately 50 kDa. In initial experiments, we cloned the Gal-A gene into a viral vector and established technical feasibility for the production and purification of this enzyme from transfected plant leaves. The yield, purity and quality of the enzyme warrant further biochemical and structural analyses to evaluate its suitability for transfusions into human subjects. PROPOSED COMMERCIAL APPLICATION: This technology will establish plants as an important and economical source of the large quantities of recombinant proteins necessary to advance modern molecular therapies. Cost advantages are based on the rapidity of the viral transfection, the favorable levels of product enrichment in crude extracts, and the high yield of product obtained from plant biomass, a renewable and low-input and easily scalable source.

成功开发负担得起的分子疗法将需要 同时生产和生物处理的全新方法 重组蛋白质。例如，代谢性储存障碍是 最常见的一组遗传异常和许多这些条件 可能可以通过补充外源性 内切酶 然而，对于任何特定的分子缺陷， 个人受到影响，因此预计治疗费用非常高 高法布里病是一种隐性的X连锁的分解代谢缺陷， 溶酶体水解酶α-半乳糖苷酶A（Ga 1-A）的活性， 这是人类第二常见的遗传性代谢储存障碍。 本提案的总体目标是确定以下方面的可行性： 经济地生产定制设计的重组Ga 1-A用于治疗 Fabry病的治疗来自人类来源的Ga 1-A具有同源二聚体 一种糖蛋白结构，亚基分子量约为50 kDa。在最初的实验中，我们将Gal-A基因克隆到病毒载体中， 并确定了生产和纯化的技术可行性 这种酶的基因。产率、纯度和 酶的质量保证进一步的生化和结构分析 以评估其对人类受试者输血的适用性。 拟定商业应用： 这项技术将使植物成为一种重要的、经济的 大量重组蛋白的来源， 推进现代分子疗法。成本优势基于 病毒转染的快速性、有利的产物水平 粗提物的富集，以及从粗提物中获得的产物的高产率， 植物生物质是一种可再生、低投入、易于扩展的资源。