Jul 14, 2014 when making the transition from conventional dishbased cell culture to bioreactors, it can be less daunting to start with smaller vessels and scale up if needed, after conditions are optimized. But if you need your cells to produce large amounts of, say, a particular secreted protein, all those dishes can be limiting not to mention cumbersome. Application of htp microfluidic culture systems to media and process optimization. Sep 14, 2017 designing a successful process to produce a particular biologic depends not only on fitting together the right cell culture media, cell line, and bioreactor, but also on careful control and. Our goal during cell culture process development is to develop high titer, scalable. Our innovative cell culture tools allow you to rapidly passage and harvest your cells using simplified, efficient workflows to scale up your optimized process. The ideal cell culture scaleup operation the objectives of cell culture scaleup are to maximize cell growth and productivity by optimizing the process parameters. Datadriven tools for optimizing cell culture media labkey. An efficient approach to cell culture medium optimization a. As discussed above, different cell lines and even cell clones require different formulations to optimize performance and the process type, that is batch.
A significant challenge for the cell and gene therapy industry is to develop a hek293t suspension cell culture process that is well characterized and can be scaledup for production whilst ensuring clinical and commercial success. The poliovirus production is usually performed in bioreactors and its production optimization is normally achieved by varying the operational parameters. It can be used for a range of different cell lines and applications. The acoustic device separates the cell suspension by a combined action of gravimetric and oscillating. Optimization of midstream cell lysis and virus filtration. In the cell culture section, serumfree lowprotein media powder is dissolved in.
How to optimize cell culture media to speed biopharma. Sartorius launched a new closedsystem product, mycap. Cellular and process bottlenecks are identified and capitalized for immediate optimization and shortterm gains in process improvements. Overcome your cell culture, purification, and process. Most cell culturists grow cell cultures in dishes, flasks or tubes.
Optimizing cell culture technology part of the ninth annual. Optimization of cell culture and cell disruption processes to enhance the production of thermophilic cellulase fncel5a in li using response surface methodology. Jiang and others published fedbatch cell culture process optimization. We offer a fully scalable cell line cellca cho, complete with cell culture media for your process and detailed product characterization biooutsource. Cell culture optimization creative biolabs provides customdesigned cell culture optimization service. We offer a fully scalable cell line cellca cho, complete with cell culture media for your process and detailed product characterization.
The use of a microbioreactor platform is, in this regard, a powerful tool for the scaling of culture conditions. Jun 27, 2018 highthroughput media and feed screening for cell culture process optimization with multiparallel micro bioreactors in this live webinar dr marta ruiz guillen, usp development specialist at 3p. This exercise shows how its done using modde doe data analytics software. Apart from the productivity of a specific cell line, the culture medium has an important impact on both yield and quantity of therapeutic protein and monoclonal antibody produced. Highthroughput media and feed screening for cell culture process optimization with multiparallel micro bioreactors in this live webinar dr marta ruiz. Results effect of early cell dilution on human t cell expansion. On day 5 of culture, the cells were transferred to 2 l xuri cellbag bioreactors.
Process optimisation for a gscho cell line 4301 3560 2829 1917 585 334 9 antibody mgl iteration 4 26 31 20 14 4 2 fold increase. Rapid development and optimization of cell culture media. Design of experiments helps optimize cell culture bioproduction system serumfree culture media used in bioprocessing can typically have 60 90 components at differing concentrations for a single cell. On day 3 of culture, fresh medium was added to increase the total culture volume by 2, 4, and 8fold. The creation of an integrated in silico model of the cell that has been tested and refined for robustness and accuracy and that is used for predicting cellular behavior in culture.
One of the talks was a perfect fit as it discussed a process that brought a product from dna to ind in a record 9month timeline for for biologics drug development in this case, recombinant protein therapeutics produced from mammalian cell culture. This doe approach to medium optimization is more efficient than the traditional linear process. Optimization of midstream cell lysis and virus filtration steps in an adenovirus purification process this application note describes screening of detergents for release of adenovirus from hek293 host cells to replace the traditionally used detergent triton x100, now on the authorization list annex xiv of registration, evaluation. Here we discuss a rationally integrated approach that best serves the practices of fedbatch cell culture process optimization. The modelling of cell culture processes has gained increasing importance in the last decades because it can be applied to design, optimize and control production processes. Futhermore, media used to grow different cell lines for bioprocessing applications may each require unique optimal chemical formulations. Culture and harvest your cells faster with more efficiency. Optimizing cell culture technology has long been considered the must attend international conference focused solely on culturing cells. Optimally in a fedbatch cell culture process, the basal medium supports cells through their growth phase and thus plays a major role in cell growth, whereas the feed medium sustains cells through the production phase for a primary contribution of increasing igg productivity.
Throughout any development process, quality by design qbd concepts are utilized. Optimization of cell culture processes for the production. The time required to customize a culture medium for a particular cell line depends upon the defining criteria, such as the starting point of the medium and the goals. Transition from medium and feed development to process optimization.
Culture and harvest your cells faster with more efficiency our innovative cell culture tools allow you to rapidly passage and harvest your cells using simplified, efficient workflows to scale up your optimized process. The media mixture layout we designed using designexpert software. But as cell culture scales up to the bioprocess level, tflasks just cant cut it, and neither do the growth conditions that were used in them. Modelassisted design of process strategies for cell culture. It also highlighted the kind of collaboration that was necessary to achieve such timeline.
The same bioreactors used for vero cell culture were used for virus infection. In academic labs, cells are grown mostly in culture dishes. Purified t cells were seeded at 1 x 10 6 cellsml in cell culture flasks and stimulated with the immunocult human cd3cd28 t cell activator. Designing a successful process to produce a particular biologic depends not only on fitting together the right cell culture media, cell line, and bioreactor, but also on careful control and. Support for cell culture media and upstream process. Basal medium, feed medium, and process parameter settings are the three major factors for fedbatch cell culture process optimization.
May 01, 20 in this study, we describe a new blending design for the medium development of a fedbatch cell culture process, enabling the optimization of all medium components of a proprietary medium in one single experiment. Biopharmaceutical process optimization with simulation and. Puzzle pieces stay the same, but process componentsmedia, cell lines, and bioreactor. Cell culture process development fujifilm diosynth. Register now and learn more about how the micromatrix offers the right solution for screening and process optimization for cell culture. Register for this miniwebinar to learn about how the mycap. Process development for protein therapeutics is increasingly dependent on highthroughput ht technologies to accelerate the screening of many conditions and the optimization of cell culture process outputs. This is in contrast with traditional 2d cell cultures in which cells are grown in a flat monolayer on a plate. A design of experiments doe approach to media optimization for chobased processes can speed things up. This leading meeting explores todays evolving strategies and technologies for improving mammalian cell cultivation, including genome engineering, crispr, modeling and computational insights and analysis in the continuing efforts to improve yield.
How to optimize cell culture media to speed biopharma development. Streamlined strategy for rapid development and optimization of cell culture media. The overall approach of optimization of medium and. Mar 25, 2018 mathematical model as key for process design. In this study, we describe a new blending design for the medium development of a fedbatch cell culture process, enabling the optimization of all medium components of a proprietary medium in one single experiment. Nowadays the implementation of scaledown methods for screening and optimization is essential in the development of new therapies. Reduction of charge variants by cho cell culture process. Quality control in biologic cell culture applications. Pdf optimization of cell culture and cell disruption. Intracellular response to process optimization and impact on. Save time and benefit from automated staining and mixing. Jan 22, 2019 these data sets on the basis of metabolic cell and process models to either optimize the conditions for a specific clone and product, or to continuously improve the production platform as a whole.
Vero cells grown on cytodex microcarrierrs using readytoprocess. Cellculture advances test bioreactor performance models. Successful optimization of chinese hamster ovary cho cell culture for biomanufacturing has a significant impact on productivity, protein quality, manufacturing efficiency and economic feasibility. Design of experiment doe approach in cell culture medium. Appropriate analyzers are critical to reliable process monitoring and control. Optimization of hek293t suspension cultivation with a doe. The cedex hires analyzer also represents the industrial standard for biopharmaceutical production. We tailor our programs to meet the needs to each program, and especially for. Chinese hamster ovary cells cho scifeed alert for new publications never miss any articles matching your research from any publisher. When basal medium optimization improved cell growth, we hypothesized that a. Sep 24, 2018 cell culture is a central component of protein development and the media used during this process can have a significant impact on its performance. The time required to customize a culture medium for a particular cell line depends upon the defin ing criteria, such as the starting point of the medium and the goals needed e.
Cell culture media are complex entities that encompass large quantities of raw materials as well as a high level of technical expertise, driving up the cost per liter. Data were exported as fcs files and analyzed using flowjo software. Mechanistic modeling and applications for cho cell culture. Cell culture to bioproduction bioprocessing summit europe. Highthroughput cell culture experimentation with bioprocessors simcell platform. An efficient approach to cell culture medium optimization. Implementation to bioprocesses using an advanced software platform. C2map is a novel cell culture media analysis platform that can carry out seamless analysis from automated sample pretreatment to simultaneous analysis of up to 95 compounds.
Mammalian cell culture manufacturers are required to test for potential mycoplasma contamination. In addition, the powerful mixture experiments coupled with the doe software provide an invaluable tool for boosting cell growth and productivity. Improvements in cell lines, expression systems and genetic engineering, and culture media and additives have resulted in greatly improved titers in recent years, and the biopharmaceutical manufacturing industry has enjoyed substantial progress in upstream manufacturing. Process optimization in upstream from cell line development to production our team will help speedup your development. In order to evaluate the process optimization results, it is crucial to calculate and compare cell specific rates and observed yields. Scaling up your cell cultures to bioreactors biocompare. As a result, researchers looking to scale up to bioreactors need to work out their culture conditions first. Cell number was corrected by bead count of flowcheck. The convenient format of cho medium optimization kit 1 allows for a rapid screening of multiple diverse cho formulations. Upstream process optimization and biomanufacturing support. Ccx cell culture expansion system that addresses the problem caused by opening flasks during cell culture expansion, that enables a production process that is less likely to become contaminated. Combined with a software platform that plots growth and product titer for each. Apr 14, 2010 cell culture process development starts with cell line generation and selection, followed by process and media optimization in small scale systems, including 96well plates, shaker flasks, and benchscale bioreactors, for high throughput screening purposes. Cell culture process development starts with cell line genera tion and selection, followed by process and media optimization in small scale systems, including 96well plates, shaker flasks, and benchscale bioreactors, for high throughput screening purposes.
The modelling of cell culture processes has gained increasing. Process optimization requires consideration of the cell type, nutrient medium, feeds, and bioreactor settings while minimizing the production of byproducts. The use of a microbioreactor platform as cell culture process. To achieve the highest performing cell culture, a fedbatch culture strategy is frequently used. An acoustic cell retention device biosep adi 1015 applisens, applikon bv, schidam, nl was used to harvest a chinese hamster ovary cho cell culture producing a recombinant protein figure 7 as an alternative to a continuous centrifuge operation. This program was commercialized in the mid1980s by batch. Process parameters such as oxygen, ph, temperature and agitation may be considered in models because of their impact on cell culture performance. Prior to virus infection, the ph, temperature and do settings were adjusted to 7. Cell culture process development starts with cell line generation and selection, followed by process and media optimization in small scale systems, including 96well plates, shaker flasks, and benchscale bioreactors, for high throughput screening purposes. In the future, predictive metabolic modeling will allow for online preemptive process optimization and control based on reduced data sets. In an effort to improve the performance of a manufacturing clone expressing a complex therapeutic protein, we investigated cell line engineering, as well as directed modulations of the cell culture process. Cell line cho igg, expressing a proprietary recombinant antibody, was transferred from a customer to sigma for custom medium development and optimization.
Dec 01, 2009 in this study, a cho cell line producing a human igg1 monoclonal antibody was subjected to a four factor, mixed level design for process optimization. The use of a microbioreactor platform as cell culture. Subject matter expertise in mammalian cell culture and related aspects such as cell metabolismmedia optimization, engineering fundamentals on cell culture doctoral degree terminal degree awarded by universities or colleges at the completion of an advanced, researchfocused course of study. Tools to optimize cell culture conditions biocompare. The vero cell culture and viral titer were monitored. We also summarize the current thinking on appropriate process development strategies and process advances that might affect process development. Cell culture processes for monoclonal antibody production. Jan zitzmann, gundula sprick, tobias weidner, christine schreiber and peter czermak may 10th 2017. So we required a combined optimization of basal and feed media to fulfill our objective. Offering new functionality, a high level of flexibility and improved performance the generation 2 system is ideally suited for applications including. A novel cell culture media analysis platform for culture. A highthroughput media design approach for high performance.
Jayanta sinha, phd, associate director, cell culture process sciences. Interested in learning how our capabilities can help enhance yours. Home insights cell culture media process optimization improvement of culture media is a common approach taken by the industry to boost upstream productivity. Cell culture process development begins with cell line generation which typically consists of a common host cell such as hek 293 or cho cells, an expression or infection system and selection, followed by process and media optimization in small scale systems such as. Software aided monitoring and control of critical process parameters helps to improve.
Sabin type 2 poliovirus was used to infect the vero cells. In this presentation, we will introduce the new streamlined strategic approach to speed media. Process optimization for recombinant protein expression in. Samples were acquired in fast mode, and volumes of 100. Reduction of charge variants by cho cell culture process optimization. Culture media the media used in this study are cho proteinfree animal componentfree medium. Biocommand software eppendorf, the vicell analyzer beckman.
Biopharmaceutical companies today are challenged to develop high producing cell lines as quickly as possible. A growth medium is designed with lower nutrient concentrations for when cell densities are low at inoculation and optimized for high specific cell growth rates to minimize culture lag phase and maintain high rates of cell growth during culture. Evaluation of a novel microbioreactor system for cell culture optimization. Introduction adenovirus advbased vectors are widely evaluated as. Using doe with micro bioreactor improves process control. On day 3, we increased the total culture volume by 2, 4, 8, 16 or 32fold, and determined the effects on total expansion. Intracellular response to process optimization and impact. Another key target is the optimization of basic process parameters such as temperature, ph, and dissolved oxygen. The ability to simultaneously test all media components presents the advantage that no potentially critical factors are missed. Kinetic model for adherent vero cell growth and poliovirus. Creative biolabs provides customdesigned cell culture optimization service. It is commonly used in gmpvalidated biopharmaceutical production processes and conforms to gmp regulations, such as 21 cfr part 11. Process optimization for recombinant protein expression in insect cells, new insights into cell culture technology, sivakumar joghi thatha gowder, intechopen, doi.
An efficient approach to cell culture medium optimization a statistical method to medium mixing. Cell culture medium optimization for upstream and scaleup processes. Screening of cho cell growing conditions upscaling of cho cell cultivations from 10 ml micromatrix to 5l stirred tank bioreactors. Optimization of cell culture processes for the production of therapeutic monoclonal antibodies. Upstream process development plus pilotscale and productionscale manufacturing support for cell culture in singleuse bioreactors.
Automated ht experimentation provides opportunities to explore a large design space by using. Optimizing cell culture technology the bioprocessing summit. Even though largescale cell culture is more of an industry than a craft. Cell culture to bioproduction bioprocessing europe. Optimization of these process parameters can be made via models for maximal cell growth and production as shown in figure 2b. The tool allows analysis and optimization of existing seed trains as well as. Performance can be improved by upgrading one or more of those three elements. A number of benchtop mini or microbioreactors are available that incorporate sensors and use software to optimize conditions. Optimization of midstream cell lysis and virus filtration steps in an.
The media used during cell culture can have a significant impact on its. Often, optimization of culture media and state variables are combined, whereas doe studies can significantly facilitate such tasks. To investigate the effect of cell density on human t cell expansion, we activated t cells on day 0 using immunocult human cd3cd28cd2 t cell activator or immunocult human cd328 t cell activator. Together we will share experiences and objectives, exploring total system integration to. C2map can provide useful information for optimization of the culture media composition and the culture process. Design of experiments with smallscale bioreactor systems. At that point, it makes sense to scale up your culture conditionsgrowing a lot more cells to make a lot more protein. To address these challenges, sartoriusstedim offers an advanced design of experiments doe approach to media optimization for chobased processes. Doe analysis using modde software and its design and analysis wizards is an important component of this process, helping to determine. Cell culture medium development is a complex process, involving the adjustment of numerous interacting components to their optimal concentrations with a goal of strong consistent support of cell growth and protein productivity. Fedbatch cell culture process optimization bioprocess. Customizing cell culture media according to individual cell line needs for each process is of paramount importance.
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