Analysis of a Novel Immobilized Cell Reactor for Unstable Recombinant Cultures

用于不稳定重组培养物的新型固定化细胞反应器的分析

• 批准号: 8710512
• 负责人: Martin Hjortso
• 金额: $ 6.74万
• 依托单位: Louisiana State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-06-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8710512&HistoricalAwards=false
• 关键词: Analysis; Novel; Immobilized; Cell; Reactor

This proposal seeks to develop an immobilized cell reactor in which a recombinant DNA instability problem (plasmid instability) is overcome by selectively killing cells without recombinant DNA (without a plasmid). This is to be done using a toxin (colicin) to E. coli cells. A plasmid is to be used that confers resistance of these cells to colicin as well as a receptor site on the cell surface to this toxin. Colicin is to be attached to a column of sepharose beads and act as a binding agent for the E. coli. Those cells without the plasmid presumably will be killed and wash off the column while those cells with the plasmid will presumably stay attached to the column. The advantage of the above system to a suspension cell culture (fermentation) is twofold. - The toxin is not in solution and will not wash out of the reactor thus saving on the cost of the toxin. - The product of the bioreactor will not need to pass through a purification step to eliminate the toxin. The PI is considered to be well qualified to carry out the proposed research and I recommend funding of this proposal.

该提案旨在开发一种固定化细胞反应器，通过选择性杀死没有重组DNA（没有质粒）的细胞来克服重组DNA不稳定问题（质粒不稳定）。 这是使用大肠杆菌细胞毒素（大肠杆菌素）来完成的。 使用质粒赋予这些细胞对大肠菌素的抗性以及细胞表面对该毒素的受体位点。 大肠杆菌素将附着在琼脂糖珠柱上，并充当大肠杆菌的结合剂。 那些没有质粒的细胞可能会被杀死并从柱上洗掉，而那些带有质粒的细胞可能会保持附着在柱上。 上述系统对于悬浮细胞培养（发酵）的优点是双重的。 - 毒素不在溶液中，不会从反应器中洗出，从而节省了毒素的成本。 - 生物反应器的产物不需要经过纯化步骤来消除毒素。 该 PI 被认为非常有资格开展拟议的研究，我建议为该提案提供资金。