Adolf Kuhner AG

Birsfelden, Switzerland

Adolf Kuhner AG

Birsfelden, Switzerland
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Raven N.,Fraunhofer Institute for Molecular Biology and Applied Ecology | Rasche S.,Fraunhofer Institute for Molecular Biology and Applied Ecology | Kuehn C.,Fraunhofer Institute for Molecular Biology and Applied Ecology | Anderlei T.,Adolf Kuhner AG | And 10 more authors.
Biotechnology and Bioengineering | Year: 2015

Tobacco BY-2 cells have emerged as a promising platform for the manufacture of biopharmaceutical proteins, offering efficient protein secretion, favourable growth characteristics and cultivation in containment under a controlled environment. The cultivation of BY-2 cells in disposable bioreactors is a useful alternative to conventional stainless steel stirred-tank reactors, and orbitally-shaken bioreactors could provide further advantages such as simple bag geometry, scalability and predictable process settings. We carried out a scale-up study, using a 200-L orbitally-shaken bioreactor holding disposable bags, and BY-2 cells producing the human monoclonal antibody M12. We found that cell growth and recombinant protein accumulation were comparable to standard shake flask cultivation, despite a 200-fold difference in cultivation volume. Final cell fresh weights of 300-387g/L and M12 yields of ∼20mg/L were achieved with both cultivation methods. Furthermore, we established an efficient downstream process for the recovery of M12 from the culture broth. The viscous spent medium prevented clarification using filtration devices, but we used expanded bed adsorption (EBA) chromatography with SP Sepharose as an alternative for the efficient capture of the M12 antibody. EBA was introduced as an initial purification step prior to protein A affinity chromatography, resulting in an overall M12 recovery of 75-85% and a purity of >95%. Our results demonstrate the suitability of orbitally-shaken bioreactors for the scaled-up cultivation of plant cell suspension cultures and provide a strategy for the efficient purification of antibodies from the BY-2 culture medium. © 2014 Wiley Periodicals, Inc.


Tissot S.,Ecole Polytechnique Federale de Lausanne | Farhat M.,Ecole Polytechnique Federale de Lausanne | Hacker D.L.,Ecole Polytechnique Federale de Lausanne | Anderlei T.,Adolf Kuhner AG | And 3 more authors.
Biochemical Engineering Journal | Year: 2010

Efficient mixing in bioreactors is essential in order to avoid concentration gradients which can be harmful for mammalian cells. To study mixing and its scalability in orbitally shaken cylindrical bioreactors, we measured mixing times in containers with nominal volumes from 2 to 1500. L with a colorimetric method using two pH indicators. Four operating parameters were tested: the liquid height, the shaking diameter, the agitation rate, and the inner diameter of the container. The mixing time decreased as the agitation rate increased until a minimal value was reached. As the shaking diameter was reduced, a higher agitation rate was needed to reach the minimal mixing time. The liquid height did not have a significant effect on the mixing time, but for a constant volume, an increase of the inner diameter slightly reduced the mixing time. The fastest mixed zones were close to the wall of the container while the zone in the center of the bulk liquid was the last to achieve homogeneity. Our study showed that the free-surface shape correlated with the mixing regime and that by keeping the inner-to-shaking diameter ratio as well as the Froude number (Fr) constant, the free-surface shapes and the mixing regimes of a 1500-L bioreactor could be mimicked in a 30-L bioreactor. We concluded that the mixing in orbitally shaken cylindrical bioreactors ensures homogeneity for mammalian cell cultures at scales up to 1500. L and that the inner-to-shaking diameter is a suitable scale-up factor for mixing. © 2010 Elsevier B.V.


Rodriguez G.,University College London | Weheliye W.,University College London | Anderlei T.,Adolf Kuhner AG | Micheletti M.,University College London | And 2 more authors.
Chemical Engineering Research and Design | Year: 2013

The mixing dynamics in a cylindrical shaken bioreactor are investigated by means of velocity and kinetic energy and mixing time measurements obtained with phase-resolved PIV and a dual pH indicator system, respectively. The objective of the work is to correlate the kinetic energy of the flow and the mixing number measured under different operating conditions. The results provide evidence that the onset of a laminar-turbulent flow transition occurs when the previously reported transition to out-of-phase flow takes place, and that the mixing number is highly dependent on the position of the feeding pipe. Insertion close to the vessel walls and thus outside the vortical structures present near the centre of the reactor can enhance mixing. © 2013 The Institution of Chemical Engineers.


Werner S.,Institute For Biotechnologie | Eibl R.,Institute For Biotechnologie | Lettenbauer C.,Sartorius Stedim Switzerland AG | Roll M.,Sartorius Stedim Switzerland AG | And 13 more authors.
Chimia | Year: 2010

Innovative mixing principles in bioreactors, for example using the rocking of a platform to induce a backwards and forwards 'wave', or using orbital shaking to generate a 'wave' that runs round in a cylindrical container, have proved to be successful for the suspension cultures of cells, especially when combined with disposable materials. This article presents an overview of the engineering characteristics when these new principles are applied in bioreactors, and case studies covering scales of operation from milliliters to 1000 liters. © Schweizerische Chemische Gesellschaft.


Rodriguez G.,University College London | Anderlei T.,Adolf Kuhner AG | Micheletti M.,University College London | Yianneskis M.,University College London | Ducci A.,University College London
Biochemical Engineering Journal | Year: 2014

Accurate determination of the mixing time in orbitally shaken bioreactors (OSRs) is essential for the optimization of mixing processes and minimization of concentration gradients that can be deleterious to cell cultures. The Dual Indicator System for Mixing Time (DISMT) was employed to measure mixing times in cylindrical and Erlenmeyer flask bioreactors. Various aspects of importance for the acquisition of accurate data from the measurement methodology are discussed, utilizing also comparisons of DISMT and pH probe results obtained in two stirred reactors. The OSR results are juxtaposed with data previously reported in the literature for both cylindrical reactors and Erlenmeyer flasks. The employment of a critical Froude number shows promise for the establishment of a scaling law for mixing time across the various types and sizes of shaken bioreactors. © 2013 Elsevier B.V.


Rodriguez G.,University College London | Pieralisi I.,University of Bologna | Anderlei T.,Adolf Kuhner AG | Ducci A.,University College London | Micheletti M.,University College London
Chemical Engineering Research and Design | Year: 2015

The flow dynamics in cylindrical shaken bioreactors of different conical bottom geometries is investigated by means of phase-resolved particle image velocimetry. Amid the variety of shaken vessels used for bioprocess applications, a cylindrical bioreactor with a conical bottom geometry was selected to assess its potential application in three-dimensional cultures, and improve solid suspension in shaken systems. This work builds upon the study of Weheliye et al. (2013. AIChE J. 59, 344) for a flat bottom with the objective to evaluate the effects of conical shaped bottoms of different heights on the fluid flow under different operating conditions with water being the working fluid. The results provide evidence that the presence of the conical bottom affects the transition from laminar to turbulent flow documented by Weheliye et al. (2013. AIChE J. 59, 344). The conical bottom with the greatest height investigated extended significantly the range of speeds over which flow transition occurs, with high intensity vortical structures spanning over the entire height of the bioreactor at lower speeds than those reported for a flat bottom geometry. This combined with the observed higher levels of kinetic energy should provide more efficient mechanisms for solid suspension. © 2015 The Institution of Chemical Engineers.


Klockner W.,RWTH Aachen | Gacem R.,RWTH Aachen | Anderlei T.,Adolf Kuhner AG | Raven N.,Fraunhofer Institute for Molecular Biology and Applied Ecology | And 3 more authors.
Journal of Biological Engineering | Year: 2013

Background: Among disposable bioreactor systems, cylindrical orbitally shaken bioreactors show important advantages. They provide a well-defined hydrodynamic flow combined with excellent mixing and oxygen transfer for mammalian and plant cell cultivations. Since there is no known universal correlation between the volumetric mass transfer coefficient for oxygen kLa and relevant operating parameters in such bioreactor systems, the aim of this current study is to experimentally determine a universal kLa correlation.Results: A Respiration Activity Monitoring System (RAMOS) was used to measure kLa values in cylindrical disposable shaken bioreactors and Buckingham's π-Theorem was applied to define a dimensionless equation for kLa. In this way, a scale- and volume-independent kLa correlation was developed and validated in bioreactors with volumes from 2 L to 200 L. The final correlation was used to calculate cultivation parameters at different scales to allow a sufficient oxygen supply of tobacco BY-2 cell suspension cultures.Conclusion: The resulting equation can be universally applied to calculate the mass transfer coefficient for any of seven relevant cultivation parameters such as the reactor diameter, the shaking frequency, the filling volume, the viscosity, the oxygen diffusion coefficient, the gravitational acceleration or the shaking diameter within an accuracy range of +/- 30%. To our knowledge, this is the first kLa correlation that has been defined and validated for the cited bioreactor system on a bench-to-pilot scale. © 2013 Klöckner et al.; licensee BioMed Central Ltd.


Grant
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: KBBE-2008-3-1-05 | Award Amount: 4.40M | Year: 2009

The CoMoFarm project will establish high-yielding production systems for pharmaceutical and industrial proteins based on plants, plant tissue and plant cells. The aim is to develop systems in which both the production host and the product itself show consistent yield and quality. The project will include a comparison of four alternative systems hydroponic plants, root cultures, moss and suspension cells, and will involve the evaluation of different species, strain and process optimization, scale-up, downstream processing, protein characterization and process evaluation in terms of regulatory compliance. As well as furthering the development of an emerging production technology, the project will incorporate numerous innovative elements such as in-process monitoring and automation of environmental parameters to maintain a consistent environment and optimize protein yield and homogeneity, novel downstream processing technologies and innovative bioreactor and hydroponic designs to maintain plant health and ensure production according to good manufacturing practice. This project will have an immense impact on the biopharmaceuticals industry, by allowing SMEs interested in molecular farming to develop plant-made pharmaceuticals without worrying about regulatory constraints or poor investment prospects. This will go a long way to establishing molecular farming as a feasible industry within Europe, and will open new revenue streams for companies currently not interested in (or aware of) molecular farming, e.g. horticulture companies and nurseries. The cost of drugs will also begin to fall, which will positively re-enforce the benefits of plants and lead to greater growth in the market.

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