Large-scale production and purification of biological compounds

生物化合物的大规模生产和纯化

• 批准号: 7593413
• 负责人: Joseph Shiloach
• 金额: $ 30.26万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7593413
• 关键词: Adsorption; Bacillus anthracis; Bacteria; Beds; Biochemistry; Biological; Biotechnology; Cell Density; Cells; Cells Plant Tissue; Commercial Sources; Corynebacterium diphtheriae; Development; Engineering; Escherichia coli; Fermentation; G-Protein-Coupled Receptors; G-substrate; Genus staphylococcus; Growth; Hela Cells; Human; Immunotoxins; Influenza; Insecta; Laboratory Research; Maintenance; Mammalian Cell; Methods; Microbiology; Molecular Biology; National Institute of Diabetes and Digestive and Kidney Diseases; Numbers; Operative Surgical Procedures; Pichia; Polysaccharides; Preparation; Process; Production; Proteins; Pseudomonas aeruginosa toxA protein; Range; Recombinant Proteins; Recombinants; Recovery; Research; Research Project Grants; Saccharomyces cerevisiae; Salmonella; Source; T-Cell Depletion; Tissues; Vaccines; Yeasts; adiponectin; cancer type; drug development; instrumentation; microorganism; structural biology

A significant portion of the Units activities is the production of high-quality and unique biologicals that are not available from commercial sources. On average, the Unit performs between 150 to 200 processes per year, which include fermentation of bacteria, yeast, insect cells, and mammalian cells (in volumes ranging from 2 to 200 liters), and recovery and purification of biomolecules, such as proteins and polysaccharides, from different sources. The following are examples of processes performed: growth of bacteria like Escherichia coli, (recombinant and native), Corynebacteria diphtheria, Salmonella, Shighella, Staphylococcus, Haefmofillus influenzae, Bacillus Anthracis, and various recombinant yeast strains like Saccharomyces cerevisiae and Pichia pastoris. In addition, mammalian cells such as HeLa, CHO, BHK and HEK 293, and insect cells like Sf 9, and High five for transient expression of recombinant proteins were propagated. Several proteins were produced and purified in gram quantities. Examples are: recombinant Pseudomonas aeruginosa exotoxin A, and human adiponectin. The various products were needed for a number of collaborative research projects: the preparation of immunotoxins for the treatment of specific types of cancer and T cell depletion, human adiponectin was made for biological studies and several proteins were produced for structural studies among them Toll proteins and G protein-coupled receptors.

该股活动的一个重要部分是生产商业来源无法获得的高质量和独特的生物制品。该单位平均每年进行150至200个过程，包括细菌、酵母、昆虫细胞和哺乳动物细胞的发酵(体积从2至200升不等)，以及从不同来源回收和纯化生物分子，如蛋白质和多糖。以下是所执行的过程的例子：细菌的生长，如大肠杆菌，(重组和天然)，白喉棒状杆菌，沙门氏菌，SHighella，葡萄球菌，流感嗜血杆菌，炭疽杆菌，以及各种重组酵母菌株，如酿酒酵母和毕赤酵母。此外，还对HeLa、CHO、BHK、HEK293等哺乳动物细胞和SF9、High Five等昆虫细胞进行了瞬时表达重组蛋白的培养。生产和纯化了几种克数量的蛋白质。例如：重组铜绿假单胞菌外毒素A和人脂联素。 许多合作研究项目需要各种产品：用于治疗特定类型癌症和T细胞耗竭的免疫毒素的制备，用于生物学研究的人脂联素，以及用于结构研究的几种蛋白质，其中包括Toll蛋白和G蛋白偶联受体。