The Experts below are selected from a list of 288 Experts worldwide ranked by ideXlab platform
Wilfried A.m. Bakker - One of the best experts on this subject based on the ideXlab platform.
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assessment of mass transfer and mixing in rigid lab scale Disposable Bioreactors at low power input levels
Biotechnology Progress, 2014Co-Authors: Gerco Van Eikenhorst, Yvonne E. Thomassen, Wilfried A.m. BakkerAbstract:Mass transfer, mixing times and power consumption were measured in rigid Disposable stirred tank Bioreactors and compared to those of a traditional glass bioreactor. The volumetric mass transfer coefficient and mixing times are usually determined at high agitation speeds in combination with sparged aeration as used for single cell suspension and most bacterial cultures. In contrast, here low agitation speeds combined with headspace aeration were applied. These settings are generally used for cultivation of mammalian cells growing adherent to microcarriers. The rigid Disposable vessels showed similar engineering characteristics compared to a traditional glass bioreactor. On the basis of the presented results appropriate settings for adherent cell culture, normally operated at a maximum power input level of 5 W m−3, can be selected. Depending on the Disposable bioreactor used, a stirrer speed ranging from 38 to 147 rpm will result in such a power input of 5 W m−3. This power input will mix the fluid to a degree of 95% in 22 ± 1 s and produce a volumetric mass transfer coefficient of 0.46 ± 0.07 h−1. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1269–1276, 2014
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Transfer of an adherent Vero cell culture method between two different rocking motion type Bioreactors with respect to cell growth and metabolic rates
Process Biochemistry, 2012Co-Authors: Yvonne E. Thomassen, Gerco Van Eikenhorst, Joyce E. Van Der Welle, Wilfried A.m. BakkerAbstract:Abstract Rapid and simple cell and virus cultivation can currently be carried out using Disposable Bioreactors. The CELL-tainer ® (CELLution Biotech BV) Disposable bioreactor is a rocking-type bioreactor which not only has vertical movement but horizontal displacement as well. Due to this two-directional movement relatively high mass-transfer capacities can be reached when compared with conventional rocking motion-type Bioreactors. Using the design of experiments (DoE) approach we have developed models for the mixing times in both the CELL-tainer ® and the BIOSTAT ® CultiBag RM (Sartorius Stedim Biotech) bioreactor (standard rocking motion-type). The conditions for cultivation of Vero cells in the CELL-tainer ® bioreactor were chosen based on comparable mixing times. Vero cells growing adherent to Cytodex 1 microcarriers were cultivated in the CELL-tainer ® and in the BIOSTAT ® CultiBag RM. Both Bioreactors were controlled with regard to temperature, pH and % dissolved oxygen. Vero cell growth in both Bioreactors was comparable with respect to the growth characteristics and main metabolite production and consumption rates. Additionally, polio virus production in both Bioreactors was shown to be similar.
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Platform Technology for Viral Vaccine Production: Comparison Between Attached and Suspension Vero Cells
Proceedings of the 21st Annual Meeting of the European Society for Animal Cell Technology (ESACT) Dublin Ireland June 7-10 2009, 2011Co-Authors: Yvonne E. Thomassen, Gerco Van Eikenhorst, Wilfried A.m. BakkerAbstract:Vero cells can be used as host for the production of many different types of viruses. The standard Vero culturing process consists of growing Vero cells adherent to microcarriers. Due to surface limitation the cells will have to be transferred to new microcarriers regularly. This subculturing of the cells, especially at large scale, is a complicated technique. This drawback can be overcome by using single cell suspension cultures. Vero cells growing in single cell suspension (sVero) were compared to adherent growing Vero cells (aVero) for their capability of producing polio virus. Using flow cytometry it was determined that sVero cells contained the polio virus receptor and were able to produce polio virus. These results indicate that sVero cells may be an ideal candidate for a platform based viral vaccine production approach. The combination of sVero cells with the flexibility of Disposable Bioreactors completes their suitability for these purposes.
Dieter Eibl - One of the best experts on this subject based on the ideXlab platform.
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Bioreactor Design and Scale-Up
Cell and Tissue Reaction Engineering, 2020Co-Authors: Gerardo Catapano, Dieter Eibl, Regine Eibl, Peter Czermak, Ralf PörtnerAbstract:Design and selection of cell culture Bioreactors are affected by cell-specific demands, engineering aspects, as well as economic and regulatory considerations. Mainly, special demands such as gentle agitation and aeration without cell damage, a well controlled environment, low levels of toxic metabolites, high cell and product concentrations, optimized medium utilization, surface for adherent cells, and scalability have to be considered. This chapter comprises engineering aspects of bioreactor systems (design, operation, scale-up) developed or adapted for cultivation of mammalian cells, such as Bioreactors for suspension culture (stirred-tank reactors, bubble columns, and air-lift reactors), fixed bed and fluidized bed reactors, hollow fiber and membrane reactors, and, finally, Disposable Bioreactors. Aspects relevant for selection of Bioreactors are discussed. Finally, an example is given of how to grow mammalian suspension cells from cryopreserved vials to laboratory and pilot scale.
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Disposable Bioreactors for cultivation of plant cell cultures
2014Co-Authors: Nicolai Lehmann, Ina Dittler, Mari Lamsa, Anneli Ritala, Heiko Rischer, Dieter Eibl, Kirsimarja Oksmancaldentey, Regine EiblAbstract:The trend for using Disposable Bioreactors in modern biotechnological processes has also been adopted for plant cell cultivations. In fact, plant cell cultures are now being grown in Disposable Bioreactors with volumes up to 400 L. This trend has been witnessed for both the development and commercial manufacture of therapeutic proteins, secondary metabolite-based pharmaceuticals and cosmetic compounds. Prominent examples of commercial products are Protalix’s ELELYSO and Mibelle Biochemistry’s Phyto Cell Tech-derived bioactive compounds.
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Disposable Bioreactors for inoculum production and protein expression
Methods of Molecular Biology, 2014Co-Authors: Regine Eibl, Christian Loffelholz, Dieter EiblAbstract:: Disposable Bioreactors have been increasingly implemented over the past ten years. This relates to both R & D and commercial manufacture, in particular, in animal cell-based processes. Among the numerous Disposable Bioreactors which are available today, wave-mixed bag Bioreactors and stirred Bioreactors are predominant. Whereas wave-mixed bag Bioreactors represent the system of choice for inoculum production, stirred systems are often preferred for protein expression. For this reason, the authors present protocols instructing the reader how to use the wave-mixed BIOSTAT CultiBag RM 20 L for inoculum production and the stirred UniVessel SU 2 L for recombinant protein production at benchtop scale. All methods described are based on a Chinese hamster ovary (CHO) suspension cell line expressing the human placental secreted alkaline phosphatase (SEAP).
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Encyclopedia of Industrial Biotechnology - Antibody Manufacture, Disposable Systems
Encyclopedia of Industrial Biotechnology, 2010Co-Authors: Regine Eibl, Dieter EiblAbstract:The growing demand for biotherapeutics and, in particular, antibodies has resulted in an increasing use of Disposables over the last ten years. This concerns both, process development and commercial manufacturing, where animal cells are grown in suspension or on microcarriers. The use of Disposable devices have been implemented in all three stages of biotechnological production processes: (i) upstreaming, (ii) downstreaming as well as (iii) final formulation and filling. The majority of applications are to be found in upstream processing. Here, Disposable expendable laboratory items, simple peripheral elements, and equipment for unit operations are well-established. Among the components listed, Disposable Bioreactors have achieved the highest growth rate over the last year. Following an introduction in which the term “single-use” is defined, a general overview of single-use devices in antibody production processes is given. We focus on frequently used Disposable bioreactor types, their characteristics, and obtainable results, and discuss apparent trends for Disposables in antibody manufacture. Keywords: characteristics; Disposable system overview; downstream processing; membrane bioreactor; orbitally shaken bioreactor; stirred Bioreactors; terminology; upstream processing; wave-mixed Bioreactors
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Disposable Bioreactors: the current state-of-the-art and recommended applications in biotechnology
Applied Microbiology and Biotechnology, 2010Co-Authors: Regine Eibl, Stephan Kaiser, Renate Lombriser, Dieter EiblAbstract:Disposable Bioreactors have increasingly been incorporated into preclinical, clinical, and production-scale biotechnological facilities over the last few years. Driven by market needs, and, in particular, by the developers and manufacturers of drugs, vaccines, and further biologicals, there has been a trend toward the use of Disposable seed Bioreactors as well as production Bioreactors. Numerous studies documenting their advantages in use have contributed to further new developments and have resulted in the availability of a multitude of Disposable bioreactor types which differ in power input, design, instrumentation, and scale of the cultivation container. In this review, the term “Disposable bioreactor” is defined, the benefits and constraints of Disposable Bioreactors are discussed, and critical phases and milestones in the development of Disposable Bioreactors are summarized. An overview of the Disposable Bioreactors that are currently commercially available is provided, and the domination of wave-mixed, orbitally shaken, and, in particular, stirred Disposable Bioreactors in animal cell-derived productions at cubic meter scale is reported. The growth of this type of reactor system is attributed to the recent availability of stirred Disposable benchtop systems such as the Mobius CellReady 3 L Bioreactor. Analysis of the data from computational fluid dynamic simulation studies and first cultivation runs confirms that this novel bioreactor system is a viable alternative to traditional cell culture Bioreactors at benchtop scale.
Regine Eibl - One of the best experts on this subject based on the ideXlab platform.
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Bioreactor Design and Scale-Up
Cell and Tissue Reaction Engineering, 2020Co-Authors: Gerardo Catapano, Dieter Eibl, Regine Eibl, Peter Czermak, Ralf PörtnerAbstract:Design and selection of cell culture Bioreactors are affected by cell-specific demands, engineering aspects, as well as economic and regulatory considerations. Mainly, special demands such as gentle agitation and aeration without cell damage, a well controlled environment, low levels of toxic metabolites, high cell and product concentrations, optimized medium utilization, surface for adherent cells, and scalability have to be considered. This chapter comprises engineering aspects of bioreactor systems (design, operation, scale-up) developed or adapted for cultivation of mammalian cells, such as Bioreactors for suspension culture (stirred-tank reactors, bubble columns, and air-lift reactors), fixed bed and fluidized bed reactors, hollow fiber and membrane reactors, and, finally, Disposable Bioreactors. Aspects relevant for selection of Bioreactors are discussed. Finally, an example is given of how to grow mammalian suspension cells from cryopreserved vials to laboratory and pilot scale.
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Disposable Bioreactors for cultivation of plant cell cultures
2014Co-Authors: Nicolai Lehmann, Ina Dittler, Mari Lamsa, Anneli Ritala, Heiko Rischer, Dieter Eibl, Kirsimarja Oksmancaldentey, Regine EiblAbstract:The trend for using Disposable Bioreactors in modern biotechnological processes has also been adopted for plant cell cultivations. In fact, plant cell cultures are now being grown in Disposable Bioreactors with volumes up to 400 L. This trend has been witnessed for both the development and commercial manufacture of therapeutic proteins, secondary metabolite-based pharmaceuticals and cosmetic compounds. Prominent examples of commercial products are Protalix’s ELELYSO and Mibelle Biochemistry’s Phyto Cell Tech-derived bioactive compounds.
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Disposable Bioreactors for inoculum production and protein expression
Methods of Molecular Biology, 2014Co-Authors: Regine Eibl, Christian Loffelholz, Dieter EiblAbstract:: Disposable Bioreactors have been increasingly implemented over the past ten years. This relates to both R & D and commercial manufacture, in particular, in animal cell-based processes. Among the numerous Disposable Bioreactors which are available today, wave-mixed bag Bioreactors and stirred Bioreactors are predominant. Whereas wave-mixed bag Bioreactors represent the system of choice for inoculum production, stirred systems are often preferred for protein expression. For this reason, the authors present protocols instructing the reader how to use the wave-mixed BIOSTAT CultiBag RM 20 L for inoculum production and the stirred UniVessel SU 2 L for recombinant protein production at benchtop scale. All methods described are based on a Chinese hamster ovary (CHO) suspension cell line expressing the human placental secreted alkaline phosphatase (SEAP).
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A New Approach for Rapid Development of Spodoptera frugiperda/BEVS-Based Processes
Proceedings of the 21st Annual Meeting of the European Society for Animal Cell Technology (ESACT) Dublin Ireland June 7-10 2009, 2011Co-Authors: Christoph Ries, Virginia Wasem, Dorothea Karrer, Corinne John, Regine EiblAbstract:The orbitally shaken CultiFlask 50 Disposable bioreactor, also known as the TubeSpin bioreactor (developed at the Ecole Polytechnique Federal, Lausanne and ExcellGene), has been used for the first time for high throughput screening to determine the optimal infection parameters (MOI: multiplicity of infection, TOI: time of infection, TOH: time of harvest) in an insect cell culture/BEVS-derived process development. Furthermore, the scale-up of the production process to Disposable shake flasks and wave-mixed CultiBags has been evaluated. With optimal infection parameters (MOI of 0.1, TOI of 1.0 × 106 cells mL–1), identical infection kinetics, similar quantities (up to 50 mg L–1) and comparable quality of the intracellular expressed catalytic domain of B-Raf kinase were obtained. The time at which the maximal average cell diameters were detected was consistently used as the TOH. It was evident from the trials that direct scale-up from CultiFlask 50 Disposable Bioreactors to wave-mixed CultiBags is possible in the case of Sf-21 cell culture/BEVS-derived processes. This new approach will contribute to savings in time and cost of 20–30 % with rapid production (approx. 4 weeks) of insect cell-derived high-value proteins in the mg range ensured.
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Encyclopedia of Industrial Biotechnology - Antibody Manufacture, Disposable Systems
Encyclopedia of Industrial Biotechnology, 2010Co-Authors: Regine Eibl, Dieter EiblAbstract:The growing demand for biotherapeutics and, in particular, antibodies has resulted in an increasing use of Disposables over the last ten years. This concerns both, process development and commercial manufacturing, where animal cells are grown in suspension or on microcarriers. The use of Disposable devices have been implemented in all three stages of biotechnological production processes: (i) upstreaming, (ii) downstreaming as well as (iii) final formulation and filling. The majority of applications are to be found in upstream processing. Here, Disposable expendable laboratory items, simple peripheral elements, and equipment for unit operations are well-established. Among the components listed, Disposable Bioreactors have achieved the highest growth rate over the last year. Following an introduction in which the term “single-use” is defined, a general overview of single-use devices in antibody production processes is given. We focus on frequently used Disposable bioreactor types, their characteristics, and obtainable results, and discuss apparent trends for Disposables in antibody manufacture. Keywords: characteristics; Disposable system overview; downstream processing; membrane bioreactor; orbitally shaken bioreactor; stirred Bioreactors; terminology; upstream processing; wave-mixed Bioreactors
Ashok Kumar - One of the best experts on this subject based on the ideXlab platform.
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Disposable polymeric cryogel bioreactor matrix for therapeutic protein production
Nature Protocols, 2013Co-Authors: Era Jain, Ashok KumarAbstract:Low cost and high efficiency make Disposable Bioreactors feasible for small-scale therapeutic development and initial clinical trials. We have developed a cryogel-based Disposable bioreactor matrix, which has been used for production of protein therapeutics such as urokinase and monoclonal antibodies (mAbs). The protocol discusses the application of a cryogel bioreactor for mAb production. Cryogels composed of either polyacrylamide (PAAm) coupled to gelatin or semi-interpenetrating PAAm-chitosan are synthesized by free-radical polymerization at −12 °C. Hybridoma cells are immobilized over the cryogel bioreactor and incubated for 48 h. Medium is circulated thereafter at 0.2 ml min^−1 and Bioreactors can be run continuously for 60 d. The cryogel-based packed-bed bioreactor can be formulated as a monolith or as beads; it also has an efficiency four times what can be obtained using a tissue-culture flask, a high surface-to-volume ratio and effective nutrient transport. After incubation, the bioreactor setup will take about 60 min using a pre-prepared sterilized cryogel.
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Upstream processes in antibody production: Evaluation of critical parameters
Biotechnology Advances, 2008Co-Authors: Era Jain, Ashok KumarAbstract:The demand for monoclonal antibody for therapeutic and diagnostic applications is rising constantly which puts up a need to bring down the cost of its production. In this context it becomes a prerequisite to improve the efficiency of the existing processes used for monoclonal antibody production. This review describes various upstream processes used for monoclonal antibody production and evaluates critical parameters and efforts which are being made to enhance the efficiency of the process. The upstream technology has tremendously been upgraded from host cells used for manufacturing to Bioreactors type and capacity. The host cells used range from microbial, mammalian to plant cells with mammalian cells dominating the scenario. Disposable Bioreactors are being promoted for small scale production due to easy adaptation to process validation and flexibility, though they are limited by the scale of production. In this respect Wave Bioreactors for suspension culture have been introduced recently. A novel bioreactor for immobilized cells is described which permits an economical and easy alternative to hollow fiber bioreactor at lab scale production. Modification of the cellular machinery to alter their metabolic characteristics has further added to robustness of cells and perks up cell specific productivity. The process parameters including feeding strategies and environmental parameters are being improved and efforts to validate them to get reproducible results are becoming a trend. Online monitoring of the process and product characterization is increasingly gaining importance. In total the advancement of upstream processes have led to the increase in volumetric productivity by 100-fold over last decade and make the monoclonal antibody production more economical and realistic option for therapeutic applications. © 2007 Elsevier Inc. All rights reserved.
Thomas Scheper - One of the best experts on this subject based on the ideXlab platform.
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differential inductive sensor for continuous non invasive cell growth monitoring in Disposable Bioreactors
2017Co-Authors: Maria Allers, Thomas Scheper, Dorte Solle, T Reinecke, Kai Bakes, Tamanna Nagraik, Marc Berger, Stefan ZimmermannAbstract:In this work, we present a low-cost sensor system for continuous non-invasive cell growth monitoring, especially for single use bioreactor (SUB) applications. The sensor system is based on a differential transformer. Using this differential setup, the influence of the primary magnetic flux is eliminated from the measuring signal, enabling highly sensitive non-invasive detection of permittivity changes in the culture medium. To evaluate the sensor, E. coli cultivations are performed and the cell density is measured through the polymer foil of a SUB. We found a linear dependency with low data scattering between measuring signal and cell density.
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sensors for Disposable Bioreactors
Engineering in Life Sciences, 2017Co-Authors: Christoph Busse, Kenneth F Reardon, Philipp Biechele, Ingo De Vries, Dorte Solle, Thomas ScheperAbstract:Modern bioprocess monitoring demands sensors that provide on-line information about the process state. In particular, sensors for monitoring bioprocesses carried out in single-use Bioreactors are needed because Disposable systems are becoming increasingly important for biotechnological applications. Requirements for the sensors used in these single-use Bioreactors are different than those used in classical reusable Bioreactors. For example, long lifetime or resistance to steam and cleaning procedures are less crucial factors, while a requirement of sensors for Disposable Bioreactors is a cost that is reasonable on a per-use basis. Here, we present an overview of current and emerging sensors for single-use Bioreactors, organized by the type of interface of the sensor systems to the bioreactor. A major focus is on non-invasive, in-situ sensors that are based on electromagnetic, semiconducting, optical, or ultrasonic measurements. In addition, new technologies like radio-frequency identification sensors or free-floating sensor spheres are presented. Notably, at this time there is no standard interface between single-use Bioreactors and the sensors discussed here. In the future, manufacturers should address this shortcoming to promote single-use bioprocess monitoring and control. This article is protected by copyright. All rights reserved
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non invasive online monitoring of cell growth in Disposable Bioreactors with a planar coil
Procedia Engineering, 2016Co-Authors: T Reinecke, Thomas Scheper, Philipp Biechele, M Frickhoffer, Stefan ZimmermannAbstract:Abstract To ensure high quality output of biotechnological processes, relevant process parameters need to be monitored. As bioprocesses are increasingly executed in single use Bioreactors, there is an increasing demand for new sensors applicable to these processes. In this work, we present a low-cost sensor system for continuous non-invasive cell growth monitoring, especially for single use bioreactor applications. The system consists of a planar coil connected to a low cost network analyzer. The coil is attached to the outside of the polymer foil of the single use bioreactor and an impedance spectrum is measured. To evaluate the sensor, E. coli cultivations are performed in a modified cultivation setup, which enables measurements through the polymer foil of a Sartorius BIOSTAT ® CultiBag RM, and additionally allows sampling of culture medium for optical density reference measurements. The resonance peak of the coil in the impedance spectrum, is observed as measure for the optical density. Regardless of the simple sensor construction, we found a good correlation between optical density and the damping ratio of the resonance peak.
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low cost sensor system for non invasive monitoring of cell growth in Disposable Bioreactors
Procedia Engineering, 2015Co-Authors: T Reinecke, Thomas Scheper, Philipp Biechele, V Schulte, Stefan ZimmermannAbstract:Abstract To ensure productivity and product quality, the parameters of biotechnological processes need to be monitored. Along temperature or pH, one important parameter is the cell density in the culture medium. In this work, we present a low-cost sensor system for online cell growth monitoring in Bioreactors via permittivity measurements based on coplanar transmission lines. To evaluate the sensor, E. coli cultivations are performed. We found a good correlation between optical density of the culture medium and the effective permittivity at a frequency of 1 kHz when the sensor is submerged into the culture medium. Measurements at higher frequencies additionally allow monitoring the osmolarity. Furthermore, an improved sensor was successfully used for first non-invasive measurements through the polymer wall of a Disposable bioreactor.
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Process analytical sensors and image-based techniques for single-use Bioreactors
Engineering in Life Sciences, 2011Co-Authors: Arne Bluma, Anne Glindkamp, Tim Höpfner, Andreas Prediger, Sascha Beutel, Thomas ScheperAbstract:In the field of biotechnology, the estimation of relevant process parameters is still a challenge. Particularly with regard to efficient control of processes with high product yields, a real-time and reliable documentation with process analytical sensors is essential. Moreover, stricter regulations of the US Food and Drug Administration for the pharmaceutical industry concerning process documentation, imposed by the Process and Analytical Technology Initiative, have increased the demand for sensors that are able to very accurately monitor and document the production process. For the monitoring and control of bioprocesses, a huge variety of sensors is used. Most systems are based on optical or electrochemical sensors. Furthermore, the market demand for Disposable sensor systems increases rapidly as the application of Disposable Bioreactors represents the biggest technical advance in modern biotechnology. These presterilized plastic bags significantly reduce the cost/income ratio and build a meaningful concept for the design and implementation of standardized peripheral bioproduction. Simple and flexible adaptation of sensors is a bottleneck and high-production requirements of the sensor components have to be met. This contribution gives an overview on concepts of sensor systems and image-based techniques that allow the online estimation of relevant process parameters used in single-use bioreactor systems.