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: 7970066
• 负责人: Joseph Shiloach
• 金额: $ 32.36万
• 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/7970066
• 关键词: Adsorption; Bacteria; Bacterial Toxins; Beds; Biochemistry; Biocompatible Materials; Biological; Biotechnology; Cell Density; Cells; Cells Plant Tissue; Clostridium; Development; Diphtheria Toxin; Engineering; Escherichia coli; Fermentation; Genus staphylococcus; Growth; Haemophilus influenzae; Hela Cells; Human; Insecta; Laboratory Research; Maintenance; Mammalian Cell; Methods; Microbiology; Molecular Biology; National Institute of Diabetes and Digestive and Kidney Diseases; Operative Surgical Procedures; Pichia; Polysaccharides; Process; Production; Protease Inhibitor; Proteins; Recombinant Proteins; Recombinants; Recovery; Research; Research Project Grants; Saccharomyces cerevisiae; Salmonella; Source; Tissues; Toxin; Vaccines; Yeasts; adiponectin; alpha 1-Antitrypsin; drug development; glycosylation; instrumentation; large scale production; microorganism; 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 between 100 to 150 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 , Bronchaseptica sp, Micobacteria sp, and various recombinant yeast strains like Saccharomyces cerevisiae and Pichia pastoris. In addition, mammalian cells such as HeLa, CHO, 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 Clostridium difficle toxin A, recombinant human adiponectin, recombinant human alpha1 proteinase inhibitor and recombinant Diphtheria toxin. The various products were needed for a number of collaborative research projects: human adiponectin was made for biological studies, human alpha 1 proteinase inhibitor for glycosylation studies, the bacterial toxins for vaccine development and several proteins were produced for structural studies among them Toll proteins.

实验室活动的很大一部分是商业来源无法获得的高质量和独特生物学的生产。平均而言，该单元每年执行100至150个过程，其中包括细菌，酵母，昆虫细胞和哺乳动物细胞的发酵（在2至200升的量中）以及来自不同来源的蛋白质和多糖类等生物分子的恢复和纯化。以下是执行过程的例子：诸如大肠杆菌（重组和天然），沙门氏菌，shighella，葡萄球菌，葡萄球菌，流感嗜血杆菌的细菌的生长，支气管菌SP，微杆菌SP，微型细菌SP，以及各种重组酵母菌菌株，如糖含量塞雷维斯和Pichiaeris和Pichiaeris。此外，传播了哺乳动物细胞，例如HELA，CHO和HEK 293，以及SF 9等昆虫细胞，而高五个用于重组蛋白的短暂表达。产生了几种蛋白质并以克数量纯化。例子是：重组艰难毒素A，重组人脂联素，重组人α1蛋白酶抑制剂和重组白喉毒素。 许多协作研究项目需要各种产品：生物脂联素是为了生物学研究，人α1蛋白酶抑制剂进行糖基化研究，用于疫苗开发的细菌毒素，并生产了几种蛋白质，用于结构研究，其中包括收集蛋白质。