Large-scale Production & Purification Of Compounds With

大规模生产

基本信息

批准号：
6503224
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
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

项目摘要

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 Saccharomyces cerevisiae, recombinant Pichia pastoris, mutant strains of Corynebacteria diphtheria, several types of Salmonella and Shigella and mutant strain of Bacillus anthracis; 2) propagating large volumes of various eucaryotic cells such as HeLa, CHO, drosophila and Sf-9; 3) purifying gram quantities of diphtheria toxin CRM (Cross Reacting Material) and recombinant Pseudomonas aeruginosa exotoxin A needed for the preparation of conjugated vaccines. In 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 performed experiments implementing expanded bed adsorption techniques for capturing proteins on ion exchangers, chelating and hydrophobic resins; 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 suppressing proteolytic activity; and 4) conducting experiments for improving the production protocol of recombinant proteins from Pichia pastoris, especially endostatin. In addition, the unit continues its research work in two topics, one is on the effect of various high density growth strategies of recombinant E.coli, on the production of recombinant proteins, and the other is 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 Salmonella typhi, and shigella flexneri. 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。该单元承担了各种各样的生物生产任务，例如原核生物和桉树的大规模生长，以及生产和纯化生物化合物，尤其是蛋白质。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：一个完全运行的多功能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，重组糖含量，重组的肉酱，重组肉芽菌，肉骨菌，肉毒杆菌和多种类型的肉芽素和肉芽素的corynenebacteria crom菌株2）传播大量各种催眠细胞，例如HeLa，Cho，果蝇和SF-9； 3）净化克量的毒素毒素CRM（交叉反应材料）和重组假单胞菌铜绿毒素Exotoxin是制备共轭疫苗所需的。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回收方法，在这里，我们进行了实施扩展的床吸附技术，以捕获离子交换器上的蛋白质，螯合和疏水性树脂； 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）通过抑制蛋白水解活性，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，尤其是内观蛋白的重组蛋白的生产方案。此外，该单元继续以两个主题继续其研究工作，一个是基于重组大肠杆菌的各种高密度生长策略，对重组蛋白的生产的影响，另一个是在Hela/vercinia系统的重组蛋白的生产过程中。该部门与LDMI，NICHD合作开发了针对Typhi沙门氏菌和Shigella Flexneri的新疫苗的工作。这些产品目前正在现场试验中。