High cell density mammalian cultures using continuous perfusion is prevalent in the biotech industry; however, as patient demand increases, continuous optimization is required to achieve higher yields while lowering cost of goods. Robust high cell density cultures must be able to maintain the health of high cell density cultures, minimize membrane fouling of cell retention devices, and maintain high harvest yields. Strategies to achieve these goals include optimization of media, evaluation of cell retention device technologies, and analysis of centrifugation or microfiltration harvest methods.

We present the first comprehensive analysis of the dynamics of Chinese hamster ovary cell-derived extracellular vesicles during the perfusion production process. The structural and elemental analysis of the fouled hollow fiber tangential flow filtration filters sheds light on hollow fiber filter fouling mechanism and strategies to mitigate filter fouling.

Many degrees of freedom are available in the operation of perfusion cell culture at scale; what is the optimal configuration accounting for Target Cell Density, VVD, duration of the perfusion run, seeding density, etc.? We have analysed this complex design space using a standard commercially available cell line as a basis. By looking at competing objectives of capital, Cost of Goods and Sustainability, a design space is identified that allows optimal configurations and an understanding of the trade-offs between economics and sustainability. We look at Process Mass Intensity and Energy Use within the facility for sustainability measures.

There is an urgent need to address the climate crisis through actionable strategies in the biopharmaceuticals sector. The significant environmental impact associated with biologics manufacturing, particularly concerning carbon emissions, has led to a focus on measuring Facilities Sustainability Performance in terms of CO2e, water use, and waste stream. However, there is an issue with Process Development's focus on Process Mass Intensity, as it does not effectively measure carbon emissions."The European Commission estimates that over 80% of all product-related environmental impacts are determined during the design phase of a product." This underscores the need to address CO2e emissions during Process Development, as manufacturing has the highest impact on the Life Cycle Footprint. We need practical tools and metrics that can be deployed to understand the carbon footprint of our processes early on in development, integrated with economic assessments to understand any trade-offs that may need to be made. In today's session, we will discuss the effectiveness of bioprocess eco-design metrics and modeling approaches that now allow us to predict and reduce the carbon footprint during early process development. We need a comprehensive framework starting in early development in the biopharmaceutical industry to achieve significant reductions in carbon emissions. This is crucial for aligning with global sustainability goals and meeting corporate expectations for net-zero transitions.

We integrated economic and ecological modeling to assess the impact of emerging technologies, including N-1 perfusion and continuous capture, on biopharmaceutical manufacturing processes. By evaluating key metrics such as cost of goods, waste generation, and energy consumption across various manufacturing scenarios, we aimed to elucidate the trade-offs and synergies between sustainability and profitability. Our findings provide valuable insights towards the development of more cost-effective and sustainable biomanufacturing processes.

As the market share of biologics continues to grow, the biopharmaceutical industry is placing increasing emphasis on sustainable production. However, quantifying the environmental impact of manufacturing processes remains a challenge. This talk will delve into existing green metrics and explore novel ones, looking at different scales, modalities, and process options. While assessing strengths and weaknesses of said metrics, opportunities for a more sustainable production are identified.

Demand for therapeutic oligonucleotides is rapidly increasing, resulting in the need for intensified downstream processes. CaptureSMB is a continuous chromatography method that has shown great promise in increasing both the capacity and productivity of chromatographic processes without increasing column size. In this presentation we describe an instance in which a CaptureSMB approach improved both the productivity and purity of a high volume, historically challenging ASO process.

mRNA-based therapeutics have emerged as cutting-edge technologies for treating various diseases. Current downstream processing, which relies on a series of chromatography methods and TFF, remains challenging with low yields and significantly impacted final production costs. We will present our integrated and continuous manufacturing process for mRNA production and purification. We are investigating novel methods for continuous mRNA precipitation-based purification, including various precipitating agents, and following precipitation with continuous flow filtration.

mRNA-LNP-based products continue to show promise, with various companies taking candidates through clinical trial. With the need for greater quantities of product, particularly for high-dose therapeutics, cost of goods and supply chain limitations may limit access to potentially revolutionary and life-saving products. This talk will discuss opportunities and challenges of mRNA process intensification, including approaches trialed at CPI with associated case data.

NGB and upstream process intensification in monoclonal cell culture have challenged platform primary recovery centrifuge and depth filtration operations with high harvest solids. NGB primary recovery changes have been evaluated such as optimization of a continuous discharge centrifuge and depth filter media to improve operational process control, improve capacity, and reduce shear. Additionally, single use harvest approaches have been demonstrated to increase flexibility. Changes have been implemented to provide a robust and capable process and have been recommended for platform updates.

We are developing an intensified continuous downstream process for monoclonal antibody (mAb) production with target precipitation for capture purification and flow-through chromatography for polish purification. The process addresses the volumetric throughput, buffer usage, and cost-of-goods bottlenecks associated with the platform Protein A-based capture step that currently limits mAb manufacturing capacity. We have processed four commercial harvest cell culture fluids and will report the corresponding process performance metrics and mAb CQAs.

Intensified upstream processes continue to exert pressure on downstream processes to efficiently purify therapeutic proteins while maintaining high product quality and ensuring removal of problematic impurities. This work focuses on the development of a polishing step for HMW removal, trace impurity clearance, and viral clearance, using a CEX resin designed specifically for flowthrough chromatography. Promising results were observed at high mass loading without sacrificing impurity removal for product related impurities and achieved 2-4 log removal of XMuLV and MVM. Resin cleaning and reuse studies were used to evaluate suitability of the resin for long-term reuse at manufacturing scale.

Regulatory authorities recommend using RTD to address material traceability in continuous manufacturing. Continuous virus filtration is an essential but poorly understood step in biologics manufacturing in respect to fluid dynamics and scale up. A model considering non-ideal mixing and film resistance for RTD is able to predict continuous virus filtration. Experimental validation using the inert tracer NaNO3 is possible. RTD influences startup and shut down of a integrated process.

Continuous manufacturing (CM) of biologics has gained significant interest in industry due to its flexibility and smaller facility footprint. Though the capital investment required by a CM suite into an existing plant is lower, optimization of the process design is important to minimize operating cost. There are also opportunities to improve the process monitoring and control strategies to ensure reliability. A hybrid approach was used to maximize productivity and fit into existing GMP facilities. The process presented is composed of 1) an intensified, high productivity continuous bioreactor process, 2) connected and automated purification steps and 3) batch downstream stages. This approach enables existing GMP facilities to accommodate a commercially viable CM process. Several case studies will be discussed with regards to process economics.

Decreasing cost of sales by increasing manufacturing productivity and lowering raw material cost is key to meet the supply for treating patients. To achieve this goal, Amgen developed an improved upstream process combining a robust new cellular host and a novel cell culture bioprocess. The increased yield from this improved process is relevant in particular for multi-specifics that can be difficult to express, emphasizing the benefit of continuously improving upstream processes. The new CHO host attained robust growth and high cell viability in both traditional fed-batch and perfusion cell culture processes, while allowing for the expression of diverse proteins from mAbs to multi-specifics, to maximize flexibility for a multi-modality pipeline. In addition, this CHO host achieved enhanced titers in an improved cell culture process through media and process optimization compared to an older CHO host. These improvements were consistently observed for different modalities at different production scales. In addition to the productivity benefits, we were able to achieve ~50% reduction in associated upstream manufacturing costs with the improved process, as well as high-throughput screening via miniaturized/automated production cultures.

In this presentation, we will discuss how our modeling shows that advanced single-use technologies deployed at the clarification and polishing unit operations can have a positive impact on key economic and sustainability metrics. We will show how using these technologies can lead to increased total output as well as reduced consumable waste, buffer consumption, lower PMI, and decreased CO₂e intensity.