Large-scale Production & Purification Of Compounds With

大规模生产

• 批准号: 6983602
• 负责人: Joseph Shiloach
• 金额: --
• 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/6983602
• 关键词: CHO cells; Corynebacterium diphtheriae; Escherichia coli; HeLa cells; Pseudomonas aeruginosa; Salmonella; Sf9 cell line; Shigella vaccines; bacterial proteins; biological products; biotechnology; chemical registry /resource; diphtheria toxin; fermentation; glucose metabolism; immunological substance; method development; microarray technology; microorganism mass culture; microorganism resource /registry; protein purification; tissue /cell culture; vaccine development

The Biotechnology Unit is responsible for the production and purification of biological material. The Unit undertakes a wide array of biological production tasks such as large scale growth of prokaryotes and eucaryotes, and production and purification of biological compounds especially proteins. These products, not available from commercial sources, are needed for clinical and structural studies. The Biotechnology Unit is capable of handling these various production tasks by implementing both physiological and technical approaches. Efficient process development and production is possible since the Unit possesses two integrated elements: a fully operational multi purpose pilot production facility and the capability of conducting research and process development. During the last year the unit performed approximately 200 different, large scale preparations including: 1) growing various microorganisms such as recombinant Escherichia coli, recombinant Pichia pastoris, mutant strains of Corynebacteria diphtheria, several types of Salmonella and Shigella, Staphylococcus strains and mutant strain of Bacillus anthracis; 2) propagating large volumes of various eucaryotic cells such as HeLa, CHO, drosophila, Sf 9 and hi five; 3) purifying gram quantities of diphtheria toxin CRM 9 (Cross Reacting Material), recombinant Pseudomonas aeruginosa exotoxin A, protective antigen from Bacillus anthracis. Staphylococcal enterotoxin type C, experimental transmission blocking vaccine for malaria and several antibodies both from hybridoma culture and rabbit serum. On the research and development arena, the unit continues its work in the following topics: 1) development of more efficient methods for large scale protein recovery, here we continue our work on implementing expanded bed adsorption techniques for capturing proteins on ion exchangers, and hydrophobic resins, utilizing new types of expanded column this column does not have flow adaptors and screens and therefore can handle larger concentration of biomass at higher low rates; 2) conducting research aimed at understanding the glucose metabolism of various recombinant E. coli strains in an effort to find out why certain strains are more suitable for foreign protein production using microarrays technology and protein analysis; 3) developing fermentation procedures for expressing sensitive, recombinant clostridium protein from E.coli by implementing anaerobic growth conditions; 4) conducting experiments for improving the production protocol of recombinant proteins from Pichia pastoris, especially experiment malaria transmission blocking vaccine; 5) the unit started another research project associated with mammalian cells, trying to understand the genetic difference between cells that can grow in suspension, and cells that are anchorage dependent. In addition, the unit continues its research work on the production process of recombinant proteins from the HeLa/vaccinia system, In collaboration with LDMI, NICHD, the unit conducted work on the development of new vaccine against Shigella flexneri and Bacillus anthracis, the products are currently in field trials.

生物技术单元负责生物材料的生产和纯化。该单元执行了广泛的生物生产任务，例如大规模的原核生物和桉树的大规模增长，以及生物化合物尤其是蛋白质的生产和纯化。这些产品（无法从商业来源获得）进行临床和结构研究。生物技术部门能够通过实施生理和技术方法来处理这些各种生产任务。由于该单元具有两个综合元素，因此有效的过程开发和生产是可能的：一个完全运行的多用途试点生产设施以及进行研究和过程开发的能力。在过去的一年中，该单元进行了大约200种不同的大规模制剂，包括：1）种植各种微生物，例如重组大肠杆菌，重组肉芽菌，巴斯德菌，二甲状腺菌的突变菌株，几种类型2）传播大量的各种念珠菌细胞，例如HeLa，Cho，Cho，果蝇，SF 9和HI五； 3）纯化的克含量毒素CRM 9（交叉反应材料），重组假单胞菌Exotoxin A，来自炭疽芽孢杆菌的保护性抗原。 C型C型肠毒素，疟疾的实验传递阻断疫苗以及杂交瘤培养和兔血清中的几种抗体。在研发领域，该单元在以下主题中继续进行工作：1）开发更有效的大规模蛋白质恢复方法，我们将继续我们的工作，在实施扩展的床床吸附技术方面，在离子交换器上捕获蛋白质，以及使用新型的列量较高的浓度，因此可以使用较高的型号来捕获离子交换器的蛋白质，并且可以使用越来越多的型树脂。 2）进行旨在了解各种重组大肠杆菌菌株的葡萄糖代谢的研究，以找出为什么使用微阵列技术和蛋白质分析的某些菌株更适合外国蛋白质生产； 3）通过实施厌氧生长条件，开发出从大肠杆菌表达敏感的重组梭菌蛋白的发酵程序； 4）进行实验，以改善Pichia Pastoris重组蛋白的生产方案，尤其是实验疟疾传播阻断疫苗； 5）该单元开始了与哺乳动物细胞相关的另一个研究项目，试图了解可以在悬浮液中生长的细胞与锚定依赖性细胞之间的遗传差异。此外，该部门继续其研究工作研究，该研究工作与HeLa/Vaccinia System的重组蛋白的生产过程，与LDMI，NICHD合作，该单元开展了针对Shhigella Flexneri和Shigella flexneri和bacillus nthracis开发新疫苗的工作，目前正在实地试验中。