Development of a novel automated perfusion mini bioreactor ‘ambr® 250 perfusion

In recent years a strong trend towards continuous biopharmaceutical processing has gathered momentum, driven by the promise of process intensification, reduced cost of goods, and more consistent and better controlled product quality. Key technologies in upstream cell culture (ATF, TFF) have enabled the start of a shift towards process intensification/continuous processing in the seed train (N-1 perfusion) and main production culture (concentrated fed-batch, perfusion) for biopharmaceutical production processes. While these technologies are now available for large scale bioreactor operations, small-scale application is limited to traditional benchtop bioreactor scales and formats. Benchtop bioreactors do provide a route to developing this new wave of intensified/continuous cell culture processes, however this approach is manually intensive, relatively low throughput and cost-intensive to operate. In the last 5 years, fed-batch cell culture process development has been significantly accelerated by wide spread implementation of the ambr 15 and ambr 250 fully automated, single-use, micro and mini bioreactor systems. Case studies will be presented on the utility of the ambr 15 as a perfusion mimic, and we also present here the first publication of a new version of the ambr 250 system currently in development ‘ambr 250 perfusion’. Technical description and operating data presented for the novel ‘ambr 250 perfusion’ system outline the capacity and capability of this technology. As established with ambr 250 for fed-batch processes, ambr 250 perfusion has the potential to provide the industry with a step change in perfusion process development capacity, enabling implementation of DoE based approaches for process optimization and characterization. It is envisaged that ‘ambr 250 perfusion’ can therefore facilitate and significantly accelerate an industry wide transition to upstream cell culture perfusion processes for novel biopharmaceuticals currently in early development

Development of a novel automated perfusion mini-bioreactor ambr® 250 perfusion

Session proposals: Towards other cell lines and systems – opportunities and challenges beyond CHO cells Pushing the Limits on Process Intensification: 10 grams/Liter and Beyond Process Scale Up/Down, Characterization and Control Strategy Definition In recent years a strong trend towards continuous biopharmaceutical processing has gathered momentum, driven by the promise of process intensification, reduced cost of goods, and more consistent and better controlled product quality. Key technologies in upstream cell culture (ATF, TFF) have enabled the start of a shift towards process intensification/continuous processing in the seed train (N-1 perfusion) and main production culture (concentrated fed-batch, perfusion) for biopharmaceutical production processes. While these technologies are now available for large scale bioreactor operations, small-scale application is limited to traditional benchtop bioreactor scales and formats. Benchtop bioreactors do provide a route to developing this new wave of intensified/continuous cell culture processes, however this approach is manually intensive, relatively low throughput and cost-intensive to operate. In the last 5 years, fed-batch cell culture process development has been significantly accelerated by wide spread implementation of the ambr 15 and ambr 250 fully automated, single-use, micro and mini bioreactor systems. Case studies will be presented on the utility of the ambr 15 as a perfusion mimic, and we also present here novel performance data of a new version of the ambr 250 system ‘ambr 250 perfusion’. Technical description and operating data and cell culture results presented for the novel ‘ambr 250 perfusion’ system outline the capacity and capability of this technology. As established with ambr 250 for fed-batch processes, ambr 250 perfusion has the potential to provide the industry with a step change in perfusion process development capacity, enabling implementation of DoE based approaches for process optimization and characterization. It is envisaged that ‘ambr 250 perfusion’ can therefore facilitate and significantly accelerate an industry wide transition to upstream cell culture perfusion processes for novel biopharmaceuticals currently in early development

From screening to process optimization: AMBR technology to speed up microbial fermentation processes

Session proposals: · Therapeutic Proteins Vaccines The development of biopharmaceuticals or biotechnological products derived from microbial fermentation is a financially risky endeavor and time consuming process, requiring technical upstream solutions which reduce timelines, increase efficiency, and raise likelihood of success. We have identified in particular the early steps of strain and process development offering best prospects to speed up the entire process significantly by using a reliable screening system. Based on the well-proven ambr® principle we designed with ambr 15 fermentation system to accelerate early stage development of microbial fermentation products. The multi-fermentation unit mimics larger scale bioreactor processes, and is suitable for screening clones, strains or growth conditions. In case studies with industrial partners using E. coli and P. pastoris, consistent and efficient control of fermentations across a variety culture conditions (e.g. feed, temperature, duration, pH) could be demonstrated. In the succeeding step of process development ambr 250 has been widely applied to speed up the 2nd critical phase of the microbial upstream process development. The larger working volume and the range of features, which this multi-parallel system offers, are superior to common benchtop fermenters. Optical density supervision, off gas analysis, fed-batch processing and advanced control capabilities allow process development for most commercial-scale upstream fermentation processes. In addition to this impressive range of features ambr250 has proven its ability to reliably increase the efficiency of fermentation process development many times through its rapid setup and cleanup, advanced control software, and automation

Developing new perfusion capabilities for ambr(R) micro and mini bioreactors

In recent years a strong trend towards intensified and continuous biopharmaceutical processing has gathered momentum, enabled by key cell culture technologies such as ATF and TFF. However, small-scale application has been limited to traditional benchtop bioreactor formats that are manually intensive, relatively low throughput and costly to operate. Automated high throughput single-use bioreactor systems have transformed fed-batch cell culture bioprocess development over the last decade and new capabilities to support perfusion culture in these micro and mini bioreactor formats could facilitate and accelerate an industry wide transition to intensified and continuous perfusion cell culture processes. Working in close collaboration with biopharm industry development partners, the design of the ‘ambr 250 high throughput’ bioreactor system has been modified to include hardware, software and single use components required to operate up to 24 parallel bioreactors with ATF or TFF cell retention modes. Iterative prototype testing with development partners has resulted in a novel ambr 250 system design capable of operating for extended culture durations and supporting high cell densities. In addition, a new Generation 2 ambr 15 cell culture system has been developed and demonstrated with technical capabilities facilitating perfusion mimic applications. Case studies will be presented on the utility of new ambr 15 system features for perfusion mimic (20-40M cell/ml) via cell settling and centrifugation methods, together with a range of industry case studies and novel performance data for the new ‘ambr 250 perfusion’ system (24 parallel perfusion cultures; \u3e30d; \u3e100M cell/ml; 0.25 vs. 5L; fully automated VCD control). As previously established with ambr systems for fed-batch processes, the new Generation 2 ambr 15 and ambr 250 perfusion systems together have the potential to provide the biopharm industry with a step change in perfusion process development capacity, enabling high throughput bioreactor screening and DoE optimization approaches for accelerated perfusion process development

Evaluation of ambr® 250 perfusion bioreactor system as a model for high-throughput perfusion cell culture process development

The efficient development and delivery of high-quality therapeutic products necessitates the need for high-throughput process development (HTPD) tools. In recent years, fed-batch process development timelines have been significantly reduced as industry has implemented fully automated mini bioreactor systems such as the ambr® 250 HT. More recently, as continuous processing for biologics has gained traction, driven by novel technology and economic pressure to significantly improve monoclonal antibody (mAb) production over the standard fed-batch process, more efficient, cost effective, and environmentally sustainable mAb processes are expected. Traditionally, perfusion process development work requires a combination of deep-well plates and small-scale stirred tank bioreactor (STR), both of which are labor intensive and time consuming. The established ambr®250 HT platform has recently been integrated with perfusion capabilities to enable rapid continuous perfusion process development (PD). In this work, we present the results from our assessment of the automated disposable perfusion bioreactor system for high throughput upstream PD activities, including clone selection and process optimization. In addition, several high stress conditions were also examined here to identify optimal operation ranges for the current system. The studies conclude that ambr® 250 perfusion reactor is able to generate process performance and product quality profiles equivalent to bench-top bioreactors for a high cell density perfusion process

Upstream microbial process characterization with single-use bioreactors from 15 mL to 50L

Developing biological and industrial molecules derived from microbial fermentation relies upon performant bioreactors to allow a rapid scale up to commercial batches. For this it is relevant to minimize any possible risks while developing a process that fits the industry quality standards. The choice of a well characterized system plays an important role from R&D through to production stages. The aim of this poster is to provide evidence to demonstrate the benefits of a microbial process developed using single-use, high throughput, and scalable upstream solutions. The method chosen to showcase this consistency is based on the DECHEMA Guidelines for Engineering Characterization principles and with the Zurich University of Applied Sciences, ZHAW. DECHEMA guidelines include a set of standard conditions for bioreactor characterization. By using process development and pilot scale bioreactors like the ambr 15f, ambr 250, and BIOSTAT STR 50, it is possible to accelerate development timelines and ensure process success

Experimental and computational fluid dynamics studies of adherent cells on microcarriers in an ambr® 250 bioreactor

Interest for microcarrier-based processes for the large-scale culture of adherent cells has recently grow, due to possible application in vaccine and cell therapy. This opportunity drives the need for effective, high-throughput, single-use, process development tools that can be translated successfully into industrial-scale systems. The automated ambr® 250 platform is one such technology, operating at a volume between 100 – 250mL, both high-throughput and single-use. The ambr250 has demonstrated significant success for suspension-based mammalian cell culture applications. However, additional investigations need to be performed on microcarrier-based processes for the culture of adherent cells. The fluid dynamics characteristics of the bioreactor must be sufficiently well understood to enable successful scale-up to larger scale bioreactors. Physical parameters such as fluid velocity, power number and shear stress are important for any cell culture. With microcarriers, there is an additional challenge as the fluid dynamics must take into account the presence of the particulate solid phase. A critical aspect for cell cultivation on microcarriers is the minimum agitator speed required to achieve complete microcarrier suspension, NJS. Under these conditions, the surface area of the attached cells is available for transfer of nutrients (including oxygen) to the cells and metabolites from them, whilst higher speeds hardly increase these transport processes and may lead to damaging fluid dynamic stresses being generated. It is also extremely beneficial to predict the flow dynamics of the stirred tank based on computational fluid dynamics (CFD). Once validated, CFD modelling is a very useful tool for analysing flow patterns, mixing time, mean and local specific energy dissipation rates, shear stress, and other parameters important for scale up in order to optimise the overall bioreactor geometry. In addition to the above fluid dynamic aspects, cell culture studies was also performed in parallel to analyse the cell growth at and around the minimum speed for microcarrier suspension, NJS. The CFD and experimental results with the single-use ambr250 bioreactor will be discussed in detail during the final presentation