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。该单位平均每年进行约100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到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)、沙门氏菌、SHighella、葡萄球菌、流感嗜血杆菌、脑膜炎奈瑟氏菌(脑膜炎球菌)、伯氏疏螺旋体、嗜盐杆菌、Bronacheptica sp、Micbacteria sp，以及各种重组酵母菌株，如巴斯德毕赤酵母。此外，对HeLa、CHO、MDCK、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、重组白喉毒素和重组疟原虫蛋白用于疟疾疫苗研究。 许多合作研究项目需要各种产品：用于生物学研究的人脂联素，用于疫苗开发的细菌毒素，以及用于结构研究的几种蛋白质，其中包括Toll蛋白质。