School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, Singapore, Singapore.
Appl Microbiol Biotechnol. 2011 Oct;92(2):241-51. doi: 10.1007/s00253-011-3513-y. Epub 2011 Aug 7.
The need to develop protein biomanufacturing platforms that can deliver proteins quickly and cost-effectively is ever more pressing. The rapid rate at which genomes can now be sequenced demands efficient protein production platforms for gene function identification. There is a continued need for the biotech industry to deliver new and more effective protein-based drugs to address new diseases. Bacterial production platforms have the advantage of high expression yields, but insoluble expression of many proteins necessitates the development of diverse and optimised refolding-based processes. Strategies employed to eliminate insoluble expression are reviewed, where it is concluded that inclusion bodies are difficult to eliminate for various reasons. Rational design of refolding systems and recipes are therefore needed to expedite production of recombinant proteins. This review article discusses efforts towards rational design of refolding systems and recipes, which can be guided by the development of refolding screening platforms that yield both qualitative and quantitative information on the progression of a given refolding process. The new opportunities presented by light scattering technologies for developing rational protein refolding buffer systems which in turn can be used to develop new process designs armed with better monitoring and controlling functionalities are discussed. The coupling of dynamic and static light scattering methodologies for incorporation into future bioprocess designs to ensure delivery of high-quality refolded proteins at faster rates is also discussed.
开发能够快速且经济有效地提供蛋白质的蛋白质生物制造平台的需求变得愈发迫切。现在，基因组的测序速度如此之快，以至于需要高效的蛋白质生产平台来进行基因功能鉴定。生物技术行业持续需要能够提供新的、更有效的基于蛋白质的药物，以应对新出现的疾病。细菌生产平台具有高表达产量的优势，但许多蛋白质的不可溶性表达需要开发多样化和优化的基于重折叠的工艺。本文综述了用于消除不可溶性表达的策略，结论是由于各种原因，包涵体难以消除。因此，需要对重折叠系统和配方进行合理设计，以加快重组蛋白的生产。本文讨论了针对重折叠系统和配方进行合理设计的努力，这些设计可以通过开发重折叠筛选平台来指导，该平台可以提供有关给定重折叠过程进展的定性和定量信息。本文还讨论了光散射技术为开发合理的蛋白质重折叠缓冲液系统带来的新机遇，这些系统反过来又可以用于开发具有更好监测和控制功能的新工艺设计。本文还讨论了将动态和静态光散射方法学结合到未来生物加工设计中，以确保以更快的速度提供高质量的重折叠蛋白质。
