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用于克隆筛选和快速缩小规模工艺优化的高通量自动化微生物生物反应器系统。

High throughput automated microbial bioreactor system used for clone selection and rapid scale-down process optimization.

作者信息

Velez-Suberbie M Lourdes, Betts John P J, Walker Kelly L, Robinson Colin, Zoro Barney, Keshavarz-Moore Eli

机构信息

The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Gower Street, Bernard Katz Building, London, WC1E 6BT, U.K.

Sartorius Stedim Biotech, York Way, Royston, Herts, SG8 5WY, U.K.

出版信息

Biotechnol Prog. 2018 Jan;34(1):58-68. doi: 10.1002/btpr.2534. Epub 2017 Aug 10.

Abstract

High throughput automated fermentation systems have become a useful tool in early bioprocess development. In this study, we investigated a 24 x 15 mL single use microbioreactor system, ambr 15f, designed for microbial culture. We compared the fed-batch growth and production capabilities of this system for two Escherichia coli strains, BL21 (DE3) and MC4100, and two industrially relevant molecules, hGH and scFv. In addition, different carbon sources were tested using bolus, linear or exponential feeding strategies, showing the capacity of the ambr 15f system to handle automated feeding. We used power per unit volume (P/V) as a scale criterion to compare the ambr 15f with 1 L stirred bioreactors which were previously scaled-up to 20 L with a different biological system, thus showing a potential 1,300 fold scale comparability in terms of both growth and product yield. By exposing the cells grown in the ambr 15f system to a level of shear expected in an industrial centrifuge, we determined that the cells are as robust as those from a bench scale bioreactor. These results provide evidence that the ambr 15f system is an efficient high throughput microbial system that can be used for strain and molecule selection as well as rapid scale-up. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:58-68, 2018.

摘要

高通量自动化发酵系统已成为早期生物工艺开发中的一种有用工具。在本研究中,我们研究了一种专为微生物培养设计的24×15 mL一次性使用微生物反应器系统ambr 15f。我们比较了该系统对两种大肠杆菌菌株BL21(DE3)和MC4100以及两种工业相关分子hGH和scFv的补料分批培养生长和生产能力。此外,还使用脉冲、线性或指数补料策略测试了不同的碳源,展示了ambr 15f系统处理自动补料的能力。我们使用单位体积功率(P/V)作为规模标准,将ambr 15f与1 L搅拌式生物反应器进行比较,该1 L搅拌式生物反应器先前已通过不同的生物系统扩大到20 L,从而在生长和产物产量方面显示出潜在的1300倍规模可比性。通过将在ambr 15f系统中生长的细胞暴露于工业离心机预期的剪切水平,我们确定这些细胞与台式生物反应器中的细胞一样健壮。这些结果证明,ambr 15f系统是一种高效的高通量微生物系统,可用于菌株和分子筛选以及快速扩大规模。©2017作者 生物技术进展 由Wiley Periodicals, Inc.代表美国化学工程师学会出版 生物技术进展,34:58 - 68,2018。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0091/5836883/40b22c3acbc7/BTPR-34-58-g002.jpg

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