Science Pool

Watch our first virtual roundtable on continuous manufacturing of biologics. Together with industry thought-leaders we aim to discuss current trends in biomanufacturing in this series. For this session we invited Brian Kelley, SVP Process and Product Development at Vir Biotechnology and Randal Bass, EVP Process Design and Biotherapeutic Operations at Just-Evotec Biologics. They discuss "How real is continuous manufacturing and what are the challenges?".

Want to learn more? Reach out to us at info@evotec.com

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Watch the 20 minute interview with Jon Gunther, VP Business Development Just-Evotec Biologics, at Biomanufacturing World Summit 2023. For a complete table of content regarding Jon's discussion, please see below.

Table of Contents

0:00 min – 01:50 min
Intro + Discussion on biomanufacturing costs and making biotherapeutics more affordable and accessible to patients worldwide

01:51 min –5:06 min
Emergence of continuous manufacturing of biologics to reduce cost of goods manufacturing (COGM)

5:07 – 7:00 min
Switching from fed-batch to continuous manufacturing, quality requirements and stance of the FDA

7:01 – 08:32 min
Timelines for switching to continuous manufacturing

08:33 –10:42 min
Pharma 4.0 / Using state-of-the-art technology incl. lights-out manufacturing approach

10:43 – 13.42 min 
Highly flexible manufacturing approach with Just-Evotec Biologics’ J.POD cGMP manufacturing facilities in North America and Europe

13.43 – 16:30 min
What should commercial partners observe in terms of cost, quality, scale-up and how this process can be de-risked

16:31 – 17:30 min
J.POD’s flexible and de-risked manufacturing approach when going from late-stage clinical to commercial supply

17:31 – 19:44
Our flexible partnership model incl. options like with our tech partnership with Sandoz for biosimilars development and commercial manufacturing

19:45 – end
Closing comments, get in touch with us!

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The biopharmaceutical industry must break with existing manufacturing paradigms if it is to reduce the cost of biological medicines and ensure they are accessible to patients around the world. The industry’s next generation of production facilities will use innovative approaches to minimize clean room space utilization, reduce the footprint of facilities and lower production costs.

In order to meet the global demand for biological medicines in these small and agile productions facilities, intensified manufacturing platforms are being developed that allow very high productivity to meet late phase clinical and commercial demand. These platforms utilize mammalian cell hosts in perfusion bioreactors linked to continuous downstream trains. Cell culture productivities have reached a point where a bioreactor can sustain a volumetric productivity of more than 2 g/L/day. This, coupled with the development of low-cost media and advancement in process automation technologies around the bioreactor, has translated to an ability for companies to operate a single-use bioreactor uninterrupted for over 15 days and execute harvest and Protein A capture on a continuous basis.

Optimizing Continuous Biomanufacturing Operating Conditions

Engineers at Just-Evotec Biologics were challenged to find the optimum operating conditions to enable the lowest production costs irrespective of facility mass output. To achieve this aim, they developed process models for our continuous process contained within a J.POD® facility. 

J.POD facilities apply modular cleanroom pod technology arranged in a controlled, non-classified ballroom to minimize the cleanroom footprint of operations that would have previously taken place in a large ballroom. Media and buffer preparations, cell expansion, upstream, downstream and post viral are all housed in separate pods. The design minimizes fixed utility infrastructure and instead relies on single-use continuous upstream and downstream operations. 

Figure 1 shows the relationship between Cost of Goods Manufactured (COGM) of a therapeutic antibody, volumetric productivity and culture duration. The results indicated that COGM is strongly inversely correlated to volumetric productivity between titers of 0.5 and 3.0 g/L/day, as shown by the sharp drop in COGM with increased volumetric productivity. The inverse correlation is not as strong when comparing COGM and culture duration, which is likely because most of the cost reduction benefits are attained when the first few kilograms of product are manufactured (e.g., because of the high cost of shared downstream disposables). Marginal increases in culture duration will generate higher amounts of product and allow the production of metric tonne-quantities of antibody but are accompanied by a proportional increase in media and downstream buffer costs.

Figure 1. Sensitivity plot illustrating Cost of Goods Manufactured (COGM) versus bioreactor design variables. Color coding was used to layer in throughput as a measure of the three variables studied.

Expanding Access to Life-Saving Medicines

Just-Evotec Biologics has two facilities in North America that are operational and manufacturing biological medicines for clients. Our European facility will be brought online in Toulouse, France in late-2024. 

These facilities in two geopolitically stable locations will provide our customers with additional supply chain security. They feature our intensified manufacturing platform allowing the agile and low-cost production of biopharmaceuticals.

The cost-modelling described in this article shows the potential for the J.POD facilities to be capable of delivering metric-tonne quantities of biological medicines for late phase clinical and commercial manufacturing. Furthermore, it illustrates how the COGM decrease dramatically as titers increase so that extremely low production costs can be achieved. We believe that this provides an opportunity to reduce the overall costs of biological medicines and will allow patients around the world greater access to these life-saving medicines.

Read The Full Article

Cell line development at Just - Evotec Biologics is focused on lowering the cost of manufacturing and creating highly productive cell lines for our clients and partners. To achieve this, we leverage our own in-house GS knockout CHO host cell line, which has been shown to support cell densities up to 80 M cell/mL and productivities upwards of 5 g/L/day in our perfusion bioreactor platform. Additionally, we have implemented advanced instruments and liquid handling automation to increase the throughput of processes that are historically major bottlenecks. Examples of this include semi-automated, independent transfections of up to 32 different expression vectors or high throughput single-cell cloning/screening of up 1000 clones. Paired with our process development and manufacturing expertise, we continue to push the envelope on improving drug costs and have enabled many of our clients to file for INDs.

If you’d like to learn more about the benefits of a customized cell line development program, you can download our white paper here

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Patients around the world have limited or restricted access to biopharmaceutical medicines. Reducing production costs while still maintaining high quality standards will help increase the affordability of biologics and ensure more patients benefit from these life-saving medicines.

Traditional biomanufacturing facilities have failed to deliver biopharmaceutical products with sufficiently low Cost of Goods Manufactured (COGM) to allow greater patient access. These facilities have been built with fixed capacity and a focus on large-scale fed-batch manufacturing. Scaling-up processes to large-scale fed-batch manufacturing facilities involves considerable risk, resource, and upfront costs. Such facilities often lack flexibility which limits the products that can be produced within them and can leave valuable production assets idle for periods of time.

Manufacturing costs link directly to capacity utilization and product demand. There is a historical precedent within the biopharmaceutical industry of operating with excess capacity but we must recognise this comes with a financial penalty. We must address this challenge if we are to respond to global healthcare emergencies, changes in the way healthcare systems are managed and greater demand for global access to biotherapeutics.


Continuous Bioprocessing Platforms and Modular Facility Designs

The industry needs new flexible biomanufacturing concepts to quickly react to market fluctuations and achieve a higher predictability of costs. Modern biopharmaceutical productions facilities use building and manufacturing technologies, such as modular construction, to minimize clean room space utilization and reduce footprints. They allow faster speed to market with a lower upfront capital investment and are readily expandable when product demand is better understood. Continuous manufacturing platforms can be integrated into these facilities for low-cost bioprocessing using mammalian cell hosts in perfusion bioreactors linked to continuous downstream trains. Production costs remain low irrespective of facility mass output, the product quality attributes are consistent and manufacturing footprints are minimized.

Just-Evotec Biologics has developed a low-cost manufacturing facility design utilizing modular cleanroom pod technology that we call J.POD®. The J.POD facility design features individual pre-fabricated cleanroom pods arranged in a controlled, non-classified ballroom to minimize the cleanroom footprint of operations that would have previously taken place in a large ballroom. Media and buffer preparations, cell expansion, upstream, downstream and post viral are all housed in separate pods. The design minimizes fixed utility infrastructure and instead relies on single-use continuous upstream and downstream operations.

Biomanufacturing Facility Cost Comparisons

We developed process models for a fed-batch process in a traditional stainless-steel facility, a fed-batch process in a single-use facility and three continuous processes in a J.POD facility. The models were created using the Biosolve software (Biopharm Software Ltd) to show the benefit of the J.POD facility design on the COGM of an antibody biologic. We used Net Present Cost (NPC) to compare scenarios. NPC estimates cash flows by computing operational costs and discounting over time using a capital parameter. It does not include revenues in the accounting of cash flows and assumes capital costs are sunk costs incurred at the beginning of the project.



Figure 1 shows the expected costs from operating the different facility types and assumes their throughput increases at a rate of 250 kg/year up until a peak value. The range selected was representative of market demands for typical biopharmaceutical manufacturing facilities. Jumps in the NPC correspond to points when the capacity of a facility is reaches and new builds are needed.

We can draw the following conclusions from the results.

1. J.POD facilities achieve remarkable production outputs despite their small footprint because of the high productivity of the continuous perfusion process.
2. Fixed CapEx comprises a large proportion of the total costs in a stainless-steel facility. J.POD facilities apply single-use technologies resulting in a shift from fixed to variable costs.
3. The fully continuous J.POD facility gives the lowest costs with the stainless-steel facility returning the highest expenditure over the range.
4. The stainless-steel facility is not being operated at its optimum utilization rate over the production output range modelled that leads to inefficiencies.

Driving Up Access to Biotherapeutic Medicines

We believe that modern biomanufacturing facilities must have smaller processing spaces, higher production throughputs and lower production costs. Modelling shows how our J.POD facilities have the lowest initial build and operating costs as well as the ability to control operating costs. These facilities are outperforming older manufacturing platforms in terms of cost and utilization. They are becoming an essential component of strategies for reducing costs and driving up access to biotherapeutic medicines. 

Read The Full Article

We recently published our latest paper in Drug Discovery Today. Our authors Fernando Garcia and Eva Gefroh introduce a type of flexible biomanufacturing facility called J.POD®, capable of capitalizing on Intensified Continuous Biomanufacturing (ICB) to deliver against current cost and speed needs in the biopharmaceutical industry. We are pleased to share this featured review with industry and partners as we continue to work together to expand access to medicines that matter.

Abstract

In this work, process models were developed to capture the impact of biomanufacturing costs on a commercial scale and emphasize the way in which facility design and operation must balance meeting product demand while minimizing production costs. Using a scenario-based modeling approach, several facility design strategies were evaluated, including a traditional large stainless-steel facility and a small footprint, portable-on-demand (POD)-based facility. Bioprocessing platforms were compared by estimating their total production costs across different facility types and specifically illustrating how continuous bioprocessing has gained in popularity as a novel and cost-effective approach to manufacture high-quality biopharmaceuticals. The analysis showed how fluctuations in market demand have a dramatic effect on manufacturing costs and plant utilization, with far-reaching implications on the total cost to patients. Read the full article in Drug Discovery Today.

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The importance of viral filtration studies

Biological therapeutics need to meet strict safety criteria. Virus safety is ensured through complementary manufacturing and quality control measures. Virus filtration is a critical element in this process, and viral filtration studies have become a key step in bioprocessing over the past decades. They are required by most regulators to bring a biologic to the market. The purpose is to assure that the final medicinal product is safe from the potential risk of viral contamination.

The quality of the design of such studies is key: when inappropriately designed, they may lead to undesired results such as non-representative filter fouling or virus breakthrough.

The characteristics of end-to-end continuous filtration

Continuous processing is a modern manufacturing method for biologicals, e.g. monoclonal antibodies (mAb). Consequently, also continuous viral clearance has to be investigated. Compared to batch filtration, continuous end-to-end processes present new challenges to the operation and validation of the viral filter:

• Higher loadings are required to maximize filter utilization and decrease filter swap outs, thereby reducing process risk;
• Higher loadings and continuous operation can lead to viral clearance assessments lasting several days and potentially overloading the filters with virus causing either non-representative filter fouling or non-representative virus breakthrough;
• Novel filter evaluation and assessment strategies are needed to maximize filter utilization while still demonstrating safety.

Consequences for assessing viral clearance in continuous end-to-end filtration

This, in turn, means that assessment of viral clearance in continuous viral filtration may require studies spanning over several days and also novel virus-spiking strategies. This type of assessment is needed to adequately demonstrate a high level of viral clearance while ensuring the filters are not overloaded with virus and are consistent with real-life manufacturing conditions.

A team of scientists at Just- Evotec Biologics has performed studies to identify a robust virus filter that retains virus despite high load challenges and low operation pressures. Key findings were:

• There are multiple filter options suitable for continuous processing;
• The use of surrogates for virus particles can give clues to filter behavior, however, the assay still needs optimization in terms load challenge.
• Alternatives to traditional virus spiking techniques such as a bracketed, integrity test approach can simplify viral clearance assessment while still demonstrating virus safety.

Summary and outlook

While bioprocessing technologies have evolved rapidly during the past decades, multiple factors such as increased cost, quality and production pressures are calling for further advances. At the same time, regulatory requirements are evolving and sophisticated safety testing has become a key prerequisite for market approval of novel biologics.

The team at JUST - Evotec Biologics has therefore kicked off an initiative to spearhead the development of next-generation bioprocessing technologies such as continuous end-to-end viral filtration.

Future work of the group will include optimization of the surrogate virus spiking and testing techniques as well as testing other virus filters for robustness of virus clearance at low flux/pressure conditions.

Interested in further information about Just-Evotec Biologics? Please visit our site.

For more information download the full poster.

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Chinese Ovary Hamster (CHO) cells are the most common mammalian cell lines used for the mass manufacturing of therapeutic proteins as they can produce recombinant proteins on the scale of 3-10 grams per liter of culture. However, the expression of these recombinant proteins rely on random genomic integration events, which typically result in a widely heterogeneous cell population. Therefore, cell line development is taking up much time for the extensive pooling and clone screening to identify clones with high expression, growth, and product quality.

Also, the random integration precludes experiments such as variant library screening as this screening method needs stably overexpressing pools or libraries of molecules in a single cell culture. With random integration, the assessment of yield, degree of library enrichment, etc. is very complicated as it is difficult to determine whether resulting library members differ due to inherent characteristics of each variant or merely to due to variable genomic integration site(s).

To address this problem and to provide a time- and cost-efficient solution, Just-Evotec Biologics’ scientists have developed two targeted integration systems by generating two clonal CHO cell lines stably expressing enhanced green fluorescent protein (eGFP) reporter landing pads in genomic hot spots. The goal of this study was to compare two options to evaluate the one that might be most suitable for in-house usage.

For the study, the team combined several approaches to circumvent random chromosomal insertions, resulting in the precision and reproducibility associated with site-specific recombinases as well as the biased selection of genomic hotspots associated with a certain transposon.

Recombination was carried out either by Cre or PhiC31 recombinase. Subsequently, genes for the expression of three therapeutic protein molecules were used to test targeted integration. The cell lines were then assayed for yield and productivity as well as characterized for landing pad copy number and integration fidelity by targeted locus amplification (TLA) and PCR. Both cell lines expressed high levels of the respective recombinant protein. The scientists additionally tested for enrichment of cell subpopulations with fully saturated landing pads with ganciclovir (GCV) counterselection.

The results of these experiments were quite compelling: Genetic characterization of the altered cell lines showed correct targeting of landing pads. Post‐integration enrichment for fully saturated landing pads using GCV counterselection increased recombinant protein titer by 2–2.5‐fold and specific productivity by ∼3.4‐fold.

Finally, the team developed a small antibody library of ~100 variants through random pairing of 10 unique light chains and 10 unique heavy chains by transfecting this library into a cell line containing a single copy landing pad wherein each cell line would express a single variant. Puromycin selection was used to identify cells that had successfully taken up one of the variants and cell-sorting for variants that successfully paired and expressed. Finally, the identity of successful chain pairs was determined with next generation sequencing.

As a result, Just-Evotec Biologics has demonstrated proof-of-principle of targeted integration systems in the CHO host cell line, with consistent genome integration into expected landing pad sites and high productivity. Moreover, test cases using three antibody or antibody-fusion therapeutic molecules showed similar levels of productivity. Finally, the team demonstrated that library screening or CHO display is feasible with the 100-member variant library. The study also reveals preliminary data from ongoing work to build upon these targeted integration systems, which includes isolating a single-copy landing pad cell line and the development of a CHO display platform.

Although additional work and optimization is still needed, the great advantage of this approach lies in the predictability with regards to the chromosomal integration of transgenes of interest. Among others, a key advantage of this approach is its ability to combine eGFP as a reporter gene and transposon-mediated integration to establish high-expression landing pad cell lines. With a significant decrease in heterogeneity between clones, it is possible to be able to reduce extensive pool screening, large scale cloning, and clone screening efforts.

All in all, defining and targeting predefined locations that promote high expression of an exogenous protein allows to develop cell lines expressing different recombinant protein therapeutics with a high degree of specificity and reproducibility - which, in turn, and will save a lot of time and costs.

For more information, please read the study or have a look at our poster.