A novel toolbox for E. coli lysis monitoring
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- 作者：Vignesh Rajamanickam ; David Wurm…
- 关键词：Bioprocess monitoring ; Data science tools ; Lysis ; Escherichia coli ; Chromatogram fingerprinting ; HPLC
- 刊名：Analytical and Bioanalytical Chemistry
- 出版年：2017
- 出版时间：January 2017
- 年：2017
- 卷：409
- 期：3
- 页码：667-671
- 全文大小：578KB
- 刊物类别：Chemistry and Materials Science
- 刊物主题：Analytical Chemistry; Biochemistry, general; Laboratory Medicine; Characterization and Evaluation of Materials; Food Science; Monitoring/Environmental Analysis;
- 出版者：Springer Berlin Heidelberg
- ISSN：1618-2650
- 卷排序：409
文摘
The bacterium Escherichia coli is a well-studied recombinant host organism with a plethora of applications in biotechnology. Highly valuable biopharmaceuticals, such as antibody fragments and growth factors, are currently being produced in E. coli. However, the high metabolic burden during recombinant protein production can lead to cell death, consequent lysis, and undesired product loss. Thus, fast and precise analyzers to monitor E. coli bioprocesses and to retrieve key process information, such as the optimal time point of harvest, are needed. However, such reliable monitoring tools are still scarce to date. In this study, we cultivated an E. coli strain producing a recombinant single-chain antibody fragment in the cytoplasm. In bioreactor cultivations, we purposely triggered cell lysis by pH ramps. We developed a novel toolbox using UV chromatograms as fingerprints and chemometric techniques to monitor these lysis events and used flow cytometry (FCM) as reference method to quantify viability offline. Summarizing, we were able to show that a novel toolbox comprising HPLC chromatogram fingerprinting and data science tools allowed the identification of E. coli lysis in a fast and reliable manner. We are convinced that this toolbox will not only facilitate E. coli bioprocess monitoring but will also allow enhanced process control in the future.