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: 8554134
• 负责人: Joseph Shiloach
• 金额: $ 63.12万
• 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/8554134
• 关键词: Adsorption; Bacteria; Bacterial Toxins; Beds; Biochemistry; Biocompatible Materials; Biological; Biotechnology; Borrelia burgdorferi; Cell Density; Cells; Cells Plant Tissue; Clostridium; Development; Diphtheria Toxin; Engineering; Escherichia coli; Exotoxins; Fermentation; Genus staphylococcus; Growth; Haemophilus influenzae; Halobacterium; Hela Cells; Human; Insecta; Laboratory Research; Maintenance; Malaria Vaccines; Mammalian Cell; Methods; Microbiology; Molecular Biology; National Institute of Diabetes and Digestive and Kidney Diseases; Neisseria meningitidis; Pichia; Plasmodium; Polysaccharides; Process; Production; Proteins; Pseudomonas; Recombinant Proteins; Recombinants; Recovery; Research; Research Project Grants; Salmonella; Source; Tissues; Toxin; Vaccine Research; Yeasts; adiponectin; drug development; halobacteria; instrumentation; large scale production; microorganism; operation; structural biology; vaccine development

A significant portion of the Labs activities is the production of high-quality and unique biologicals that are not available from commercial sources. On average, the Unit performs around 100 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), Salmonella, Shighella, Staphylococcus, Haemophilus influenzae, N. meningitidis (meningococcus), Borrelia burgdorferi, Halobacteria, Bronchaseptica sp, Micobacteria sp, and various recombinant yeast strains like Pichia pastoris. In addition, mammalian cells such as HeLa, CHO, MDCK and HEK 293, and insect cell such 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 Clostridium difficle toxin A, recombinant human adiponectin, recombinant pseudomonas aeruginasa exotoxin A, recombinant Diphtheria toxin and recombinant Plasmodium proteins for malaria vaccine research. The various products were needed for a number of collaborative research projects: human adiponectin was made for biological studies, the bacterial toxins for vaccine development and several proteins were produced for structural studies among them Toll proteins.

实验室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。该部门平均每年执行约 100 个流程，其中包括细菌、酵母、昆虫细胞和哺乳动物细胞的发酵（体积从 2 升到 200 升不等），以及从不同来源回收和纯化生物分子，例如蛋白质和多糖。以下是所执行过程的示例：大肠杆菌（重组和天然）、沙门氏菌、沙门氏菌、葡萄球菌、流感嗜血杆菌、脑膜炎奈瑟菌（脑膜炎球菌）、伯氏疏螺旋体、盐杆菌、支气管败血病菌、微杆菌和各种重组酵母菌株等细菌的生长 像毕赤酵母一样。此外，还增殖了用于瞬时表达重组蛋白的哺乳动物细胞（例如HeLa、CHO、MDCK和HEK 293）和昆虫细胞（例如Sf 9和High 5）。几种蛋白质的生产和纯化以克为单位。例如：重组艰难梭菌毒素 A、重组人脂联素、重组铜绿假单胞菌外毒素 A、重组白喉毒素和用于疟疾疫苗研究的重组疟原虫蛋白。 许多合作研究项目都需要各种产品：人类脂联素用于生物学研究，细菌毒素用于疫苗开发，多种蛋白质用于结构研究，其中包括 Toll 蛋白。