Collaborative Research: Efficient Bioseparation by Intertwining Strain, Chromatography, and Affinity Tail Design
合作研究:通过交织菌株、色谱和亲和尾部设计实现高效生物分离
基本信息
- 批准号:0533949
- 负责人:
- 金额:--
- 依托单位:
- 依托单位国家:美国
- 项目类别:Continuing Grant
- 财政年份:2006
- 资助国家:美国
- 起止时间:2006-03-01 至 2010-08-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Ataai0533949This proposal is a collaborative study with the University of Arkansas (BES 0534836) which describes a new protein optimization purification methodology via the technique of immobilized metal affinity chromatography (IMAC). If successful, the work will reduce columncapacity loss due to contaminating protein adsorption, and reduce the complexity of protein elution protocols. The work involves defining the low background region of the IMAC elution profile of E.coli proteins using both pH and alternately imidazole gradients, identifying these proteins using a combination of 2D gel electrophoresis, mass spectrometry, and database analysis, deleting the genes encoding proteins if they are unnecessary for growth under laboratoryconditions, (or altering them via specific mutation such that they no longer elute in the low protein background region of the host elution profile), and developing highly efficient metal binding peptides to direct elution towards the low background region.The methodology used to develop improvements for IMAC is broadly applicable to all affinity systems employing affinity tails. Therefore, the proposed work will catalyze a general improvement strategy for bioseparation. In addition, the students working on this project will become broadly trained in the tools of biochemical engineering as well as molecular biology thus contributing to a generation of interdisciplinary engineers and scientists capable of biotechnology leadership.
该提案是与阿肯色大学(BES 0534836)的合作研究,描述了一种新的通过固定化金属亲和层析(IMAC)技术的蛋白质优化纯化方法。如果成功,这项工作将减少由于污染蛋白质吸附而造成的柱容量损失,并降低蛋白质洗脱方案的复杂性。这项工作包括使用pH值和交替咪唑梯度来定义大肠杆菌蛋白的IMAC洗脱谱的低背景区域,使用2D凝胶电泳、质谱和数据库分析的组合来鉴定这些蛋白质,删除编码蛋白质的基因,如果它们在实验室条件下生长是不必要的。(或通过特定的突变改变它们,使它们不再在宿主洗脱谱的低蛋白背景区洗脱),并开发高效的金属结合肽,以直接洗脱低背景区。用于开发IMAC改进的方法广泛适用于所有使用关联尾的关联系统。因此,所提出的工作将催化生物分离的一般改进策略。此外,参与该项目的学生将在生化工程和分子生物学的工具方面得到广泛的培训,从而为一代跨学科的工程师和科学家做出贡献,他们有能力领导生物技术。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Mohammad Ataai其他文献
Mohammad Ataai的其他文献
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