Use of Operating Windows in the Assessment of Integrated Robotic Systems for the Measurement of Bioprocess Kinetics

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• 作者：Anthony J. Nealon ; Karen E. Willson ; Samuel C. R. Pickering ; Tim M. Clayton ; Ronan D. O'Kennedy ; Nigel J. Titchener-Hooker ; and Gary J. Lye
• 刊名：Biotechnology Progress
• 出版年：2005
• 出版时间：February 2005
• 年：2005
• 卷：21
• 期：1
• 页码：283 - 291
• 全文大小：197K
• 年卷期：v.21,no.1(February 2005)
• ISSN：1520-6033

文摘

This study examines the utility of an automated liquid handling robot integrated witha microwell plate reader to enable the rapid acquisition of bioprocess kinetic data.The relationship between the key parameters for liquid handling accuracy andprecision and the sample detection period has been characterized for typical low-viscosity (<2.0 mPa·s) aqueous and organic phases and for a high-viscosity aqueousphase (60 mPa·s), all exhibiting Newtonian rheology. The use of a simple graphicalmethod enables the suitability of a given automation platform to be assessed once theuser has determined the minimum sample detection period and the minimum accurateand precise dispense volume. This provides for a reduction in the duration of anyexperiment by maximizing well usage within each microwell plate. The suitability ofemploying an integrated automation platform to gather kinetic data for systems typicalof those encountered in bioprocessing is analyzed via a series of case studies.Application to alkaline cell lysis, where disruption is complete within 120 s, showedthat the range of available dispense volumes and the number of wells that can beutilized is limited. In contrast, analysis of a system exhibiting slow process kinetics,the fermentation of Escherichia coli TOP10 pQR239 in microwell plates, demonstratedthat, for a typical sample detection period of 30 min, the only restrictions on the degreeof well utilization are the liquid handling accuracy and precision and the volumecapacity of the liquid handling robot. Finally, liquid-liquid extraction, an example ofa kinetically independent operation, was also examined. In this case, only a singleequilibrium measurement is required, which means that the only restrictions to theutilization of the integrated devices are the liquid handling accuracy and precision.Integrated automation platforms represent a powerful process development tool overtraditional experimental methods used for bioprocess development. Smaller volumesof reagent and sample can be used to achieve greater throughput, while high levels ofreproducibility and sensitivity are maintained.