The Experts below are selected from a list of 19236 Experts worldwide ranked by ideXlab platform
Klaus Joeris - One of the best experts on this subject based on the ideXlab platform.
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automated and enhanced clone screening using a fully automated microtiter plate based system for Suspension Cell Culture
Biotechnology Progress, 2019Co-Authors: Sven Markert, Carsten Musmann, Peter Hulsmann, Klaus JoerisAbstract:Recently, we established an automated microtiter plate (MTP)-based system for Suspension Cell Culture for high-throughput (HT) applications in biopharmaceutical process development. In the present report, the new system was evaluated regarding its potential to improve clone screening by allowing high-throughput fed-batch cultivation at an early stage. For this purpose, a fully automated procedure was compared to a mainly batch mode-based manual standard process. The new system performed daily measurements of viable Cell density and product concentration for a total of 96 clones in biological duplicates that were evaluated for final clone selection. This resulted in a more than fivefold increase in sample throughput and 4 weeks of time saving compared to the reference process. The top clone characterized by the highest Cell specific productivity was identified only by the new process. In contrast, this clone was lost in the expansion phase of the reference procedure. Overall, the new system identified more high-productive clones, offering more alternatives and flexibility for process development. In-process monitoring of glucose and lactate levels representing crucial secondary selection criteria further enhanced top clone identification. Clone characterization at an early stage was further extended by linking the MTP-based Cell Culture system to additional HT-analytic systems for N-glycosylation analysis as well as gene expression analysis by reverse transcriptase-quantitative polymerase chain reaction. These powerful tools connected to the automated MTP-based Cell Culture system lead to considerably advanced quality and speed of clone screening, and increase the probability of selecting the most suitable clone. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2760, 2019.
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establishment of a fully automated microtiter plate based system for Suspension Cell Culture and its application for enhanced process optimization
Biotechnology and Bioengineering, 2017Co-Authors: Sven Markert, Klaus JoerisAbstract:We developed an automated microtiter plate (MTP)-based system for Suspension Cell Culture to meet the increased demands for miniaturized high throughput applications in biopharmaceutical process development. The generic system is based on off-the-shelf commercial laboratory automation equipment and is able to utilize MTPs of different configurations (6-24 wells per plate) in orbital shaken mode. The shaking conditions were optimized by Computational Fluid Dynamics simulations. The fully automated system handles plate transport, seeding and feeding of Cells, daily sampling, and preparation of analytical assays. The integration of all required analytical instrumentation into the system enables a hands-off operation which prevents bottlenecks in sample processing. The modular set-up makes the system flexible and adaptable for a continuous extension of analytical parameters and add-on components. The system proved suitable as screening tool for process development by verifying the comparability of results for the MTP-based system and bioreactors regarding profiles of viable Cell density, lactate, and product concentration of CHO Cell lines. These studies confirmed that 6 well MTPs as well as 24 deepwell MTPs were predictive for a scale up to a 1000 L stirred tank reactor (scale factor 1:200,000). Applying the established Cell Culture system for automated media blend screening in late stage development, a 22% increase in product yield was achieved in comparison to the reference process. The predicted product increase was subsequently confirmed in 2 L bioreactors. Thus, we demonstrated the feasibility of the automated MTP-based Cell Culture system for enhanced screening and optimization applications in process development and identified further application areas such as process robustness. The system offers a great potential to accelerate time-to-market for new biopharmaceuticals. Biotechnol. Bioeng. 2017;114: 113-121. © 2016 Wiley Periodicals, Inc.
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development and application of an automated multiwell plate based screening system for Suspension Cell Culture
BMC Proceedings, 2013Co-Authors: Sven Markert, Carsten Musmann, Klaus JoerisAbstract:The already presented automated, multiwell plate (MWP) based screening system for Suspension Cell Culture is now routinely used in process development. It is characterized by a fully automated workflow with integrated analytical instrumentation and uses shaken 6-24 well plates as bioreactors which can be run in batch and fed-batch mode with a capacity of up to 384 reactors in parallel [1]. A wide ranging analytical portfolio is available to monitor Cell Culture processes and to characterize product quality. Assays running on the screening system comprise the determination of Cell concentration and viability, quantification of nutrients and metabolites as well as detection of apoptosis level and staining of organelles. Additionally a RT-qPCR method has been setup to measure gene expression level in a high throughput manner. Having a large network in-house to high throughput groups of the analytical department a lot of advanced methods can easily be performed like chromatographic and mass spectrometry to characterize product quality. Current work focuses on expanding the analytical portfolio to develop control strategies for automated Cell Culture processes. Besides setting up a robust method for pH measurement we evaluate different spectroscopic techniques like Raman, infrared or 2D fluorescence as fast and powerful analytical tools.
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development of an automated multiwell plate based screening system for Suspension Cell Culture
BMC Proceedings, 2011Co-Authors: Sven Markert, Klaus JoerisAbstract:The automation of Cell Culture processes becomes more important in the pharmaceutical industry due to compressed timelines and the need to develop more products more efficiently. This drive to develop new processes faster and more efficient requires a streamlined workflow. Resource intensive approaches like the use of shake flasks limit the accessible design space for the development of highly productive processes or the characterization of established processes. Process automation provides the appropriate tools to address the following key points: Increasing experimental throughput ⇨ “Design of Experiments” using full factorial designs Increasing process information ⇨ improve process understanding (“Quality by Design”) Automate repetitive manual work ⇨ gain efficiency, focus on high value tasks Improve reproducibility ⇨ ensure robust processes A robotic plate handler based system was selected to meet the demands of a flexible, fast and modular screening system as presented in figure figure11. Figure 1 Schematic illustration of the developed robotic screening system prototype. Only the core system is shown with a robotic plate handler as key device connecting shaken cultivation, processing and analytical components. Results Scale-up prediction The comparability of results obtained with this new multiwell plate based Culture system for Suspension adapted Cell lines plates and bioreactors had to be verified. It could be shown that 6-well plates were predictive for a scale-up to a 1000 L stirred tank reactor. The parameter profiles of viable Cell density, lactate and antibody concentration were comparable in multiwell plates and bioreactors (2 L, 10 L and 1000 L). The plates can be used for process scale-up prediction.
Sven Markert - One of the best experts on this subject based on the ideXlab platform.
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automated and enhanced clone screening using a fully automated microtiter plate based system for Suspension Cell Culture
Biotechnology Progress, 2019Co-Authors: Sven Markert, Carsten Musmann, Peter Hulsmann, Klaus JoerisAbstract:Recently, we established an automated microtiter plate (MTP)-based system for Suspension Cell Culture for high-throughput (HT) applications in biopharmaceutical process development. In the present report, the new system was evaluated regarding its potential to improve clone screening by allowing high-throughput fed-batch cultivation at an early stage. For this purpose, a fully automated procedure was compared to a mainly batch mode-based manual standard process. The new system performed daily measurements of viable Cell density and product concentration for a total of 96 clones in biological duplicates that were evaluated for final clone selection. This resulted in a more than fivefold increase in sample throughput and 4 weeks of time saving compared to the reference process. The top clone characterized by the highest Cell specific productivity was identified only by the new process. In contrast, this clone was lost in the expansion phase of the reference procedure. Overall, the new system identified more high-productive clones, offering more alternatives and flexibility for process development. In-process monitoring of glucose and lactate levels representing crucial secondary selection criteria further enhanced top clone identification. Clone characterization at an early stage was further extended by linking the MTP-based Cell Culture system to additional HT-analytic systems for N-glycosylation analysis as well as gene expression analysis by reverse transcriptase-quantitative polymerase chain reaction. These powerful tools connected to the automated MTP-based Cell Culture system lead to considerably advanced quality and speed of clone screening, and increase the probability of selecting the most suitable clone. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2760, 2019.
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establishment of a fully automated microtiter plate based system for Suspension Cell Culture and its application for enhanced process optimization
Biotechnology and Bioengineering, 2017Co-Authors: Sven Markert, Klaus JoerisAbstract:We developed an automated microtiter plate (MTP)-based system for Suspension Cell Culture to meet the increased demands for miniaturized high throughput applications in biopharmaceutical process development. The generic system is based on off-the-shelf commercial laboratory automation equipment and is able to utilize MTPs of different configurations (6-24 wells per plate) in orbital shaken mode. The shaking conditions were optimized by Computational Fluid Dynamics simulations. The fully automated system handles plate transport, seeding and feeding of Cells, daily sampling, and preparation of analytical assays. The integration of all required analytical instrumentation into the system enables a hands-off operation which prevents bottlenecks in sample processing. The modular set-up makes the system flexible and adaptable for a continuous extension of analytical parameters and add-on components. The system proved suitable as screening tool for process development by verifying the comparability of results for the MTP-based system and bioreactors regarding profiles of viable Cell density, lactate, and product concentration of CHO Cell lines. These studies confirmed that 6 well MTPs as well as 24 deepwell MTPs were predictive for a scale up to a 1000 L stirred tank reactor (scale factor 1:200,000). Applying the established Cell Culture system for automated media blend screening in late stage development, a 22% increase in product yield was achieved in comparison to the reference process. The predicted product increase was subsequently confirmed in 2 L bioreactors. Thus, we demonstrated the feasibility of the automated MTP-based Cell Culture system for enhanced screening and optimization applications in process development and identified further application areas such as process robustness. The system offers a great potential to accelerate time-to-market for new biopharmaceuticals. Biotechnol. Bioeng. 2017;114: 113-121. © 2016 Wiley Periodicals, Inc.
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development and application of an automated multiwell plate based screening system for Suspension Cell Culture
BMC Proceedings, 2013Co-Authors: Sven Markert, Carsten Musmann, Klaus JoerisAbstract:The already presented automated, multiwell plate (MWP) based screening system for Suspension Cell Culture is now routinely used in process development. It is characterized by a fully automated workflow with integrated analytical instrumentation and uses shaken 6-24 well plates as bioreactors which can be run in batch and fed-batch mode with a capacity of up to 384 reactors in parallel [1]. A wide ranging analytical portfolio is available to monitor Cell Culture processes and to characterize product quality. Assays running on the screening system comprise the determination of Cell concentration and viability, quantification of nutrients and metabolites as well as detection of apoptosis level and staining of organelles. Additionally a RT-qPCR method has been setup to measure gene expression level in a high throughput manner. Having a large network in-house to high throughput groups of the analytical department a lot of advanced methods can easily be performed like chromatographic and mass spectrometry to characterize product quality. Current work focuses on expanding the analytical portfolio to develop control strategies for automated Cell Culture processes. Besides setting up a robust method for pH measurement we evaluate different spectroscopic techniques like Raman, infrared or 2D fluorescence as fast and powerful analytical tools.
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development of an automated multiwell plate based screening system for Suspension Cell Culture
BMC Proceedings, 2011Co-Authors: Sven Markert, Klaus JoerisAbstract:The automation of Cell Culture processes becomes more important in the pharmaceutical industry due to compressed timelines and the need to develop more products more efficiently. This drive to develop new processes faster and more efficient requires a streamlined workflow. Resource intensive approaches like the use of shake flasks limit the accessible design space for the development of highly productive processes or the characterization of established processes. Process automation provides the appropriate tools to address the following key points: Increasing experimental throughput ⇨ “Design of Experiments” using full factorial designs Increasing process information ⇨ improve process understanding (“Quality by Design”) Automate repetitive manual work ⇨ gain efficiency, focus on high value tasks Improve reproducibility ⇨ ensure robust processes A robotic plate handler based system was selected to meet the demands of a flexible, fast and modular screening system as presented in figure figure11. Figure 1 Schematic illustration of the developed robotic screening system prototype. Only the core system is shown with a robotic plate handler as key device connecting shaken cultivation, processing and analytical components. Results Scale-up prediction The comparability of results obtained with this new multiwell plate based Culture system for Suspension adapted Cell lines plates and bioreactors had to be verified. It could be shown that 6-well plates were predictive for a scale-up to a 1000 L stirred tank reactor. The parameter profiles of viable Cell density, lactate and antibody concentration were comparable in multiwell plates and bioreactors (2 L, 10 L and 1000 L). The plates can be used for process scale-up prediction.
Mohammad Reza Mofid - One of the best experts on this subject based on the ideXlab platform.
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Optimization of the basal medium for improving production and secretion of taxanes from Suspension Cell Culture of Taxus baccata L
DARU Journal of Pharmaceutical Sciences, 2012Co-Authors: Abolghasem Abbasi Kajani, Sharareh Moghim, Mohammad Reza MofidAbstract:Background and purpose of the studyTaxol is one of the most effective anticancer drugs that isolated from Taxus sp. due to the slow growth of Taxus trees and low concentration of Taxol in the tissues, the biotechnological approaches especially plant Cell Culture have been considered to produce Taxol in commercial scale.MethodsWe investigated the effects of basal medium type used in Culture media on production of Taxol and other taxane compounds from Cell Suspension Culture of T. baccata L. Briefly, five commonly basal media including Gamborg, Murashige and Skoog, Woody Plant, Schenk and Hildebrandt, and Driver and Kuniyuki medium were used for preparing separate Suspension Culture media. The intra- and extra-Cellular yields of taxanes were analyzed by using HPLC after 21 days period of culturing.ResultsThe yields of taxanes were significantly different for the Cultures prepared by different basal media. Moreover, the effects of basal medium on the yield of products differed for varius taxane compounds. Maximum yields of Baccatin III (10.03 mgl^-1) and 10-deacetyl baccatin III (4.2 mgl^-1) were achieved from the DKW basal media, but the yield of Taxol was maximum (16.58 mgl^-1) in the WPM basal media. Furthermore, the secretion of taxanes from the Cells into medium was also considerably affected by the type of basal medium. The maximum extra-Cellular yield of Taxol (7.81 mgl^-1), Baccatin III (5.0 mgl^-1), and 10-deacetyl baccatin III (1.45 mgl^-1) were also obtained by using DKW basal medium that were significantly higher than those obtained from other Culture media.
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optimization of the basal medium for improving production and secretion of taxanes from Suspension Cell Culture of taxus baccata l
DARU, 2012Co-Authors: Abolghasem Abbasi Kajani, Sharareh Moghim, Mohammad Reza MofidAbstract:Background and purpose of the study Taxol is one of the most effective anticancer drugs that isolated from Taxus sp. due to the slow growth of Taxus trees and low concentration of Taxol in the tissues, the biotechnological approaches especially plant Cell Culture have been considered to produce Taxol in commercial scale.
Neil Emans - One of the best experts on this subject based on the ideXlab platform.
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Molecular farming of pharmaceutical proteins.
Transgenic research, 2000Co-Authors: R. Fischer, Neil EmansAbstract:Molecular farming is the production of pharmaceutically important and commercially valuable proteins in plants. Its purpose is to provide a safe and inexpensive means for the mass production of recombinant pharmaceutical proteins. Complex mammalian proteins can be produced in transformed plants or transformed plant Suspension Cells. Plants are suitable for the production of pharmaceutical proteins on a field scale because the expressed proteins are functional and almost indistinguishable from their mammalian counterparts. The breadth of therapeutic proteins produced by plants range from interleukins to recombinant antibodies. Molecular farming in plants has the potential to provide virtually unlimited quantities of recombinant proteins for use as diagnostic and therapeutic tools in health care and the life sciences. Plants produce a large amount of biomass and protein production can be increased using plant Suspension Cell Culture in fermenters, or by the propagation of stably transformed plant lines in the field. Transgenic plants can also produce organs rich in a recombinant protein for its long-term storage. This demonstrates the promise of using transgenic plants as bioreactors for the molecular farming of recombinant therapeutics, including vaccines, diagnostics, such as recombinant antibodies, plasma proteins, cytokines and growth factors.
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Antibody production by molecular farming in plants
Journal of Biological Regulators and Homeostatic Agents, 2000Co-Authors: R. Fischer, Stefan Schillberg, K. Hoffmann, Neil EmansAbstract:"Molecular farming" is the production of pharmaceutical proteins in transgenic plants and has great potential for the production of therapeutic anti-cancer antibodies and recombinant therapeutic proteins. Plants make fully functional recombinant human or animal antibodies. Cultivating transgenic plants on an agricultural scale will produce almost unlimited supplies of recombinant proteins for uses in medicine. Combinatorial library technology is a key tool for the generation and optimisation of therapeutic antibodies ahead of their expression in plants. Optimised antibody expression can be rapidly verified using transient expression assays in plants before creation of transgenic Suspension Cells or plant lines. SubCellular targeting signals that increase expression levels and optimise protein stability can be identified and exploited using transient expression to create high expresser plant lines. When high expresser lines have been selected, the final step is the development of efficient purification methods to retrieve functional antibody. Antibody production on an industrial scale is then possible using plant Suspension Cell Culture in fermenters, or by the propagation of stably transformed plant lines in the field. Recombinant proteins can be produced either in whole plants or in seeds and tubers, which can be used for the long-term storage of both the protein and its production system. The review will discuss these developments and how we are moving toward the molecular farming of therapeutic antibodies becoming an economic and clinical reality.
Carsten Musmann - One of the best experts on this subject based on the ideXlab platform.
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automated and enhanced clone screening using a fully automated microtiter plate based system for Suspension Cell Culture
Biotechnology Progress, 2019Co-Authors: Sven Markert, Carsten Musmann, Peter Hulsmann, Klaus JoerisAbstract:Recently, we established an automated microtiter plate (MTP)-based system for Suspension Cell Culture for high-throughput (HT) applications in biopharmaceutical process development. In the present report, the new system was evaluated regarding its potential to improve clone screening by allowing high-throughput fed-batch cultivation at an early stage. For this purpose, a fully automated procedure was compared to a mainly batch mode-based manual standard process. The new system performed daily measurements of viable Cell density and product concentration for a total of 96 clones in biological duplicates that were evaluated for final clone selection. This resulted in a more than fivefold increase in sample throughput and 4 weeks of time saving compared to the reference process. The top clone characterized by the highest Cell specific productivity was identified only by the new process. In contrast, this clone was lost in the expansion phase of the reference procedure. Overall, the new system identified more high-productive clones, offering more alternatives and flexibility for process development. In-process monitoring of glucose and lactate levels representing crucial secondary selection criteria further enhanced top clone identification. Clone characterization at an early stage was further extended by linking the MTP-based Cell Culture system to additional HT-analytic systems for N-glycosylation analysis as well as gene expression analysis by reverse transcriptase-quantitative polymerase chain reaction. These powerful tools connected to the automated MTP-based Cell Culture system lead to considerably advanced quality and speed of clone screening, and increase the probability of selecting the most suitable clone. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2760, 2019.
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development and application of an automated multiwell plate based screening system for Suspension Cell Culture
BMC Proceedings, 2013Co-Authors: Sven Markert, Carsten Musmann, Klaus JoerisAbstract:The already presented automated, multiwell plate (MWP) based screening system for Suspension Cell Culture is now routinely used in process development. It is characterized by a fully automated workflow with integrated analytical instrumentation and uses shaken 6-24 well plates as bioreactors which can be run in batch and fed-batch mode with a capacity of up to 384 reactors in parallel [1]. A wide ranging analytical portfolio is available to monitor Cell Culture processes and to characterize product quality. Assays running on the screening system comprise the determination of Cell concentration and viability, quantification of nutrients and metabolites as well as detection of apoptosis level and staining of organelles. Additionally a RT-qPCR method has been setup to measure gene expression level in a high throughput manner. Having a large network in-house to high throughput groups of the analytical department a lot of advanced methods can easily be performed like chromatographic and mass spectrometry to characterize product quality. Current work focuses on expanding the analytical portfolio to develop control strategies for automated Cell Culture processes. Besides setting up a robust method for pH measurement we evaluate different spectroscopic techniques like Raman, infrared or 2D fluorescence as fast and powerful analytical tools.
