Intensified purification and refolding of inclusion body protein through multi-column simulated moving bed chromatography
多柱模拟移动床层析强化纯化和复性包涵体蛋白
基本信息
- 批准号:406956-2011
- 负责人:
- 金额:$ 8.08万
- 依托单位:
- 依托单位国家:加拿大
- 项目类别:Research Tools and Instruments - Category 1 (<$150,000)
- 财政年份:2010
- 资助国家:加拿大
- 起止时间:2010-01-01 至 2011-12-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Protein refolding is essential to impart biological activity on proteins which affects therapeutic efficacy. Challenges in protein refolding include low yield, high product dilution, and large process volumes, which translates to exorbitant product costs. Protein purification using single chromatographic columns are commonly used, but disadvantaged by low media utilization, high solvent consumption, and low productivity. Multi-column Continuous Chromatography (MCC) processing based on Simulated Moving Bed (SMB) technology is a promising technology that has had significant impact on pharmaceutical chiral separations, but has not found widespread use in bio-processing due to challenges in processing. This research aims to intensify refolding and purification of few commercially-relevant proteins, for improved process efficiency and productivity via MCC. The new knowledge generated from this fundamental study will address technical challenges that have caused SMB to be largely unexplored for large-scale bio-processing. Particularly, this will enable innovative rational design and scale-up methodology of MCC process aimed to enhance productivity of continuous protein bio-processing. The successful outcome of this work will have broad application for high-throughput of new biologics entering clinical trials reducing drug manufacturing costs and the speed with which new protein drugs can be brought to market. It is expected to have immediate applicability for both new protein-based drugs and "bio-generics", which are competing with drugs coming off-patent on a cost basis. The PI has a unique MCC experimental rig capable be undertaking the proposed research. The requested analytical infrastructure of Waters UPLC H-Class Bio systems will help perform reliable high-resolution rapid analysis of extremely small amounts of target molecules as diverse as intact proteins, protein complexes, and peptides upstream of mass spectrometry in LC-MS workflows. This initiative is unique as there is no similar MCC development on-going anywhere in Canada, and therefore, this investment will be creating a distinctive state-of-the-art research capability on large-scale protein refolding and purification within this country.
蛋白质重折叠对于赋予蛋白质生物活性是必不可少的,其影响治疗功效。蛋白质重折叠的挑战包括低产量、高产品稀释度和大的工艺体积,这转化为过高的产品成本。使用单一色谱柱的蛋白质纯化是常用的,但缺点是介质利用率低、溶剂消耗高和生产率低。基于模拟移动床(SMB)技术的多柱连续色谱(MCC)处理是一种很有前途的技术,其对药物手性分离产生了显著影响,但由于处理中的挑战而没有在生物处理中得到广泛应用。本研究旨在通过MCC加强少数商业相关蛋白的复性和纯化,以提高工艺效率和生产率。从这项基础研究中产生的新知识将解决导致SMB在大规模生物加工中基本上未被探索的技术挑战。特别是,这将使创新的合理设计和规模扩大的方法,MCC过程,旨在提高生产率的连续蛋白质生物加工。这项工作的成功结果将广泛应用于进入临床试验的新生物制剂的高通量,降低药物制造成本和新蛋白质药物推向市场的速度。预计它将立即适用于新的蛋白质药物和“生物仿制药”,这是竞争的药物在成本基础上的专利。PI有一个独特的MCC实验装置,能够进行拟议的研究。沃茨UPLC H-Class Bio系统所需的分析基础设施将有助于在LC-MS工作流程中对质谱上游的各种目标分子(如完整蛋白质、蛋白质复合物和肽)进行可靠的高分辨率快速分析。这一举措是独一无二的,因为在加拿大任何地方都没有类似的MCC开发,因此,这项投资将在该国建立一个独特的大规模蛋白质重折叠和纯化的最先进的研究能力。
项目成果
期刊论文数量(0)
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Ray, AjayKumar其他文献
Ray, AjayKumar的其他文献
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