Abstract
Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the throughput of such studies; however, there are limitations with their use. One important constraint is the limited number of offline samples that can be taken compared to those taken for monitoring cultures in large‐scale bioreactors. The small volume of miniature bioreactor cultures (15 mL) is incompatible with the large sample volume (600 µL) required for bioanalysers routinely used. Spectroscopy technologies may be used to resolve this limitation. The purpose of this study was to compare the use of NIR, Raman, and 2D‐fluorescence to measure multiple analytes simultaneously in volumes suitable for daily monitoring of a miniature bioreactor system. A novel design‐of‐experiment approach is described that utilizes previously analyzed cell culture supernatant to assess metabolite concentrations under various conditions while providing optimal coverage of the desired design space. Multivariate data analysis techniques were used to develop predictive models. Model performance was compared to determine which technology is more suitable for this application. 2D‐fluorescence could more accurately measure ammonium concentration (RMSECV 0.031 g L−1) than Raman and NIR. Raman spectroscopy, however, was more robust at measuring lactate and glucose concentrations (RMSECV 1.11 and 0.92 g L−1, respectively) than the other two techniques. The findings suggest that Raman spectroscopy is more suited for this application than NIR and 2D‐fluorescence. The implementation of Raman spectroscopy increases at‐line measuring capabilities, enabling daily monitoring of key cell culture components within miniature bioreactor cultures. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:337–346, 2017
Highlights
The industry in recent years has become increasingly interested in the use of spectroscopy technologies such as Raman, near-infrared (NIR), and 2D-fluorescence (2D-F) for the measuring, monitoring, and control of bioprocesses
This is, in part, a result of the American Food and Drug Administration’s (FDA) guidance on process analytical technology (PAT), which encourages the use of innovative tools and technologies to increase understanding and control of manufacturing processes.[1]
The applicability of spectroscopic methods for miniature bioreactor (MB) cultures has been reviewed[11]; the authors are not aware of any literature experimentally comparing the ability of spectroscopy techniques to monitor multiple analytes under suitable operational constraints required for daily monitoring of MB cultures
Summary
The industry in recent years has become increasingly interested in the use of spectroscopy technologies such as Raman, near-infrared (NIR), and 2D-fluorescence (2D-F) for the measuring, monitoring, and control of bioprocesses. The ICH Q8 guideline defines QbD as “a systematic approach to development that begins with predefined objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management.”[2] Spectroscopy technologies offer a nondestructive, rapid, and robust method for generating multianalyte data.[3] These technologies are used routinely to monitor chemical pharmaceutical production processes, but their application to mammalian cell culture for production of biologics is much more challenging This is primarily due to the complex matrix background[4] and low concentrations of the metabolites of interest. The applicability of spectroscopic methods for miniature bioreactor (MB) cultures has been reviewed[11]; the authors are not aware of any literature experimentally comparing the ability of spectroscopy techniques to monitor multiple analytes under suitable operational constraints required for daily monitoring of MB cultures
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