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.

生物技术股负责生物材料的生产和提纯。该单位承担广泛的生物生产任务，如原核生物和真核生物的大规模生长，以及生物化合物特别是蛋白质的生产和纯化。这些产品无法从商业来源获得，需要用于临床和结构研究。生物技术股能够通过实施生理和技术方法来处理这些不同的生产任务。高效率的工艺开发和生产是可能的，因为该股拥有两个综合要素：一个全面运作的多用途试验生产设施和进行研究和工艺开发的能力。在过去的一年中，该单位进行了大约200种不同的大规模制备，包括：1）培养各种微生物，如重组大肠杆菌、重组巴斯德毕赤酵母、白喉棒状杆菌的突变株、几种沙门氏菌和志贺氏菌、葡萄球菌菌株和炭疽芽孢杆菌的突变株; 2）大量繁殖各种真核细胞，如HeLa、CHO、果蝇、Sf 9和hi five; 3）纯化克量白喉毒素CRM 9，（交叉反应材料），重组铜绿假单胞菌外毒素A，炭疽杆菌保护性抗原。C型葡萄球菌肠毒素、实验性疟疾传播阻断疫苗以及来自杂交瘤培养物和兔血清的几种抗体。在研究和发展竞技场方面，该股继续在以下专题开展工作：1）开发用于大规模蛋白质回收的更有效的方法，在这里，我们继续我们的工作，实施用于在离子交换剂和疏水树脂上捕获蛋白质的膨胀床吸附技术，利用新型膨胀柱，该柱不具有流动适配器和筛网，费率; 2）进行旨在了解各种重组E.大肠杆菌菌株，试图找出为什么某些菌株更适合于使用微阵列技术和蛋白质分析生产外源蛋白质; 3）开发通过实施厌氧生长条件从大肠杆菌表达敏感的重组梭菌蛋白质的发酵程序; 4）进行实验以改进来自巴斯德毕赤酵母的重组蛋白的生产方案，特别是实验疟疾传播阻断疫苗; 5）该单位开始另一项与哺乳动物细胞有关的研究项目，试图了解可悬浮生长的细胞与依赖锚定的细胞之间的遗传差异。此外，该单位继续其对HeLa/牛痘系统重组蛋白生产工艺的研究工作，该单位与Linguistics，NICHD合作，开展了针对福氏志贺菌和炭疽杆菌的新疫苗开发工作，产品目前正在进行田间试验。