Optimization is key to achieving a highproducing cho cell culture. A robust base medium must first be identified and proven in small spinners or shaker cultures. In this publication, we show that at elevated pco 2 in batch and fed. Statistical methods in media optimization for batch and fed. Biocell pilot series bioreactors 60 l to 300 l dci. Automated dynamic fedbatch process and media optimization. Monoclonal antibody production was identified as being partially growth associated. Elevated pco2 affects the lactate metabolic shift in cho cell. Jan 23, 2007 hybridoma 8f producing antif monoclonal antibodies mab were grown in batch and fed batch mode with glutamine as the limiting substrate. Towards the development of automated fedbatch cell culture. Versatile modeling and optimization of fed batch processes. During cell culture process development, the main goal is to achieve high titers and a defined product quality of the recombinant molecule. Modelling batch and fedbatch mammalian cell cultures for.
Jiang and others published fedbatch cell culture process optimization. The cell densities achieved in perfusion culture 30100 x 10 6 cellsml are typically higher than for fedbatch modes 525 x 10 6 cellsml. Cell culture processes for monoclonal antibody production. Optimize fedbatch culture early in process development to save time and resources. An igg1producing cho cell line was cultured in hyclone. A road map for efficient fedbatch cell culture optimization. Gibco bioproduction services utilizes a custom design of experiment doe approach which leverages a variety of high throughput tools such as simcell and clonepix to accelerate process and cell line development. This work demonstrates the development of a fed batch process for a chinese hamster ovary cho cell line producing a monoclonal antibody mab. Media optimization for cho fedbatch processes using a doe.
These systems represent the best tool for optimization of your batch, fed batch, or continuous cell culture processes using suspended cells or microcarriers. Optimized cell culture media vital to process development. This was done automatically through the micromatrix software and required. Blends were tested on both the cell expansion and the fedbatch production phases, ensuring that. General attempts to model fed batch processes have been described for an overview see. Experimental evaluation in two independent fed batch cultures resulted in a. Automated dynamic fedbatch process and media optimization for. Optimization of bioreactor growth conditions and feeding strategy has the potential for several fold increase in heparosan yield, as a fedbatch process provides a feed medium that prevents depletion of nutrients and sustains the production phase 34.
Developing an automatically controlled feeding process in an. Sixteen media formulations were designed from 43 of 47 components. Employing the cumulative cell hour concept, external metabolic flux. Effects of feeding strategy on cho cell performance in fed. For subsequent projects involving chok1sv, fmte was added to efb or efc to help improve cell culture performance. High throughput automated micro bioreactor for high density fed batch strain selection ambr 15 fermentation is a high throughput automated bioreactor system for 24 parallel microbial fermentations in a costeffective 8 12 ml micro bioreactor format. Developing a feeding strategy a comprehensive process is required when developing a feeding strategy. Automated dynamic fedbatch process and media optimization for high productivity cell culture process. To achieve the highest performing cell culture, a fed batch culture strategy is frequently used. Cell culture optimization would be so much easier if a universal feed existed. We automated that feed using eppendorfs biocommand batch control supervisory control and data acquisition scada software.
Nov 29, 2016 fed batch culture is currently the most common industrial process for cho cell culture. Here we discuss a rationally integrated approach that best serves the practices of fedbatch cell culture process optimization. In a fedbatch process, a basal medium supports initial growth and production, and a feed medium prevents depletion of nutrients and sustains the production phase 9. I ended the conversation by asking andreas if he had anything else to add for listeners.
This approach increased inoculation cell densities and decreased cultivation times by. Automated dynamic fed batch process and media optimization for high productivity cell culture process development franklin lu,1 poh choo toh,2 iain burnett,1 feng li,1 terry hudson,1 ashraf amanullah,1 jincai li1 1oceanside pharma technical development, genentech, inc. This can be achieved through cell line engineering, media optimization and bioprocess development huang et al. We recognize that all bioprocess systems are unique. Fourteen process parameters were optimized medium and feed components and physical parameters. While the description of product concentration in the cell mass is rather. Fed batch culture is, in the broadest sense, defined as an operational technique in biotechnological processes where one or more nutrients substrates are fed supplied to the bioreactor during cultivation and in which the product s remain in the bioreactor until the end of the run. Basal medium, feed medium, and process parameter settings are the three major factors for fed batch cell culture process optimization. Cell culture process development for fedbatch using a novel. Mammalian cell culture process development clone evaluation media development process optimization screen and select the highest producing and most stable clone develop optimal growth and production media for each cell line optimize conditions for biomanufacturing process in a scaledown version scale up system.
Nov 22, 2011 initial experiments were performed using chemically defined culture medium in 125 ml shake flasks with 50 ml working volume. This process optimization toolbox will increase the titer of your cellcacho production cell line by saving 2 weeks of turnaround time, since the doe is defined and ready to go. 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. The main goal of the study was to test the feasibility of the ga approach and not to optimize the particular processes as such. In its absence, says bd, try a feed that works across multiple base media and cells in a platform process. The use of a microbioreactor platform as cell culture process. Actipro cell culture production medium is intended to be used in combination with cell boost 7a and 7b supplements to enhance recombinant protein production in fed batch or perfusion processes. Fedbatch cell culture process optimization bioprocess. He said that when optimizing fed batch culture, the manufacturing process should be kept in mind at all times. In the table the fedbatch operation is divided into two basic models, one without feedback control and the other with feedback control. A highthroughput media design approach for high performance.
Dec 01, 2009 each culture was run in a day fedbatch process with ph, do, and glucose control. 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. Utilizing its expertise in process vessel manufacturing, cell culture and automated systems, dcibiolafitte introduces its biocell pilot series, a new generation of sip cip pilot bioreactor. 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. A fedbatch operation is that operation in which one or more nutrients are added continuously or intermittently to the initial medium after the start of cultivation or from the halfway point through the batch process. Modeling of the fed batch process and optimization of qp was used to. Optimization of fedbatch processes for two recombinant mammalian cell lines was carried out using simple ga. Maintaining good culture for cells is very important in the last phase of the culture for this reason. Use the analysis wizard to model the three responses. Performance can be improved by upgrading one or more of those three elements.
How to optimize cell culture media to speed biopharma development. Applying this highthroughput method on a fed batch, rather than on a simple batch, process opens new perspectives for medium and feed development that enables identification of an optimized process in a short time frame. The 12 different fed batch designs were developed and approved as optimal solutions for a variety of different cellcacho production cell lines. Rapid optimization of fedbatch processes using the micromatrix. Optimization of process conditions for mammalian fedbatch. Details of fedbatch operation are summarized in table 1. May 01, 20 in this study, we describe a new blending design for the medium development of a fed batch cell culture process, enabling the optimization of all medium components of a proprietary medium in one single experiment. In the case of fed batch culture, the concentrated components of the nutrient are gradually added to the batch culture. Article automated dynamic fed batch process and media optimization for high productivity cell culture process development franklin lu,1 poh choo toh,2 iain burnett,1 feng li,1 terry hudson,1. This means that the fed nutrients must be supplemented in the right ratios and in sufficient amounts.
Mar 25, 2018 mathematical model as key for process design. Create a modde project and reproduce the experimental design that was used. Cell culture process development on chos cells was initially conducted with cho cd efficientfeed a efa, efb, and efc. The 12 different fedbatch designs were developed and approved as optimal solutions for a.
In the batch culture, the microorganisms are inoculated in fixed volume of medium. Selection of chemically defined media for cho cell fedbatch. 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. In this article, we discuss a strategy to improve operational procedures in an escherichia coli fed batch fermentation through automated process control. In the perfusion batch culture, the addition of the culture and withdrawal of an equal volume of used cell free medium is performed. An economic comparison of three cell culture techniques. Fedbatch biofuel production process using a new brunswick. The media mixture layout we designed using designexpert software. The ambr15 and modde software used here provided a path to obtain effective media formulations for a given specific cho cell line. Improvement and simplification of fedbatch bioprocesses with. Medium and feed optimization for fedbatch production of a. The culture broth is harvested usually only at the end of the operational period. Fedbatch is an easy and popular way to intensify cell cultivations.
How to optimize cell culture media to speed biopharma. Dynamic optimization of hybridoma growth in a fedbatch. Designing feed strategies for fedbatch cho cultures. An alternative description of the method is that of a. The design and optimization of a fermentation process plays a key role in achieving high productivity and robustness at the production scale. Basal medium, feed medium, and process parameter settings are the three major factors for fedbatch cell culture process optimization. Highthroughput media blending method for the medium development of a fed batch cell culture process. Versatile modeling and optimization of fed batch processes for the.
Nov 30, 2017 the aim of this study was to investigate the effects of elevated pco 2 concentrations on the lactate metabolic shift phenomena in cho cell culture processes. In the fedbatch process, we fed the culture starting on day 3 with cd efficientfeed c agt nutrient supplement thermo fisher scientific at 3% of the working volume per day. We here demonstrate a novel feeding strategy which is designed to hold a particular metabolic state in a fedbatch process by adaptive feeding in. Cell culture bioreactors microbial bioreactors software apps for bioreactors. Statistical methods in media optimization for batch and fedbatch animal cell culture article in bioprocess and biosystems engineering 302. Cell line stability is another factor that should be considered since volumetric and specific productivity decline as cell age increases for. Apr 14, 2010 to date, typical production cell line specific productivities range from 20 pgcellday to 70 pgcellday and peak viability cell densities from 530.
Fedbatch operation an overview sciencedirect topics. Cellretention systems add a level of complexity to the process. Based on these modeling approaches, optimization of fed batch processes has been attempted using pontryagins maximum principle 2, 3, greens theorem, or dynamic programming. The ultimate goal of this study is to establish a comprehensive dynamic model which may be used for modelbased optimization of the cell culture for mab production in both batch and fed batch systems. For the production of mabs, high performing fed batch processes have. So we required a combined optimization of basal and feed media to fulfill our objective. Cell culture media are complex entities that encompass large quantities of raw materials as well as a high level of. Media blending following a custommade mixture doe resulted in 376 different blends which were evaluated in 96dwp for cell culture performance in fed batch. Comparing culture methods in monoclonal antibody production.
Doe to accelerate the optimization of fedbatch strategies 21. Genedata bioprocess makes the development of new or optimized fedbatch and perfusion upstream processes more efficient by supporting all workflows. Finally, a multivariate analysis enabled identification of individual potential critical components for further optimization. The ability to simultaneously test all media components presents the advantage that no potentially critical factors are missed. Sep 18, 2019 impact of dynamic online fedbatch strategies on metabolism, productivity and nglycosylation quality in cho cell cultures.
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