The outsourcing market can often be subject to the trends and whims of the greater pharma market. Consider, for instance, messenger RNA (mRNA) vaccines—following the success of both Moderna and Pfizer-BioNTech’s COVID-19 vaccines, mRNA vaccines have become a “new” technology of interest, 50 plus years of prior research notwithstanding (1). Numerous companies have thrown their hat into this ring, exploring mRNA as an avenue for future vaccines for anything from zika virus to cancer (2).

However, while there is significant buzz around mRNA technologies, the companies investigating this technology remain just a fraction of the larger biopharma market. In spite of this, these few companies may be having an outsized impact on the outsourcing industry, particularly among companies specializing in biologics, or those which claim to offer “one-stop-shop” services. mRNA technologies have unique manufacturing needs that need to be incorporated into facility design for commercial scale production (3). As a result, contract manufacturing and development organizations (CDMOs) and contract research organizations (CROs) aiming to capture this market may have to turn existing plans on a dime to satisfy the needs of these potential partners, ultimately breeding a culture of adaptability among outsourcing companies who strive to capture emerging markets.

Put simply, if the pharma market is the X axis, or independent variable, then the outsourcing market is the Y axis, or dependent variable. Consequently, if one can brush past the buzzwords and marketing speak, investigating where companies are turning to outsourcing can help illuminate trends in the undercurrent of the larger biopharma space.

Before launching into integrated CDMOs, it’s important to get a firm footing on where outsourcing currently stands. According to John McDermott, vice-president of Scientific Consulting at Quotient Sciences, a pharmaceutical CDMO and CRO, historically pharmaceutical companies have favored outsourcing clinical activity and been less willing to outsource non-clinical activities like R&D, manufacturing, and commercialization.

“Outsourcing of clinical activity has been prominent in the pharmaceutical industry for a number of years, but it has taken longer for other drug development disciplines to outsource to the same extent,” says McDermott. “The reasons for this are clear; maintaining internal clinical capability carries a high fixed cost and invariably offers limited access to the wide range of patient groups required by the indications under investigation today, and so clinical absolutely rely on the outsourcing strategies. Pharma has found it easier to maintain internal resources in other disciplines, such as formulation development and clinical trial manufacturing, and therefore has been slower to fully outsource.”

However, according to McDermott, the emergence of biotechs and specialist formulation technology for solubility enhancement or even modified release has increased outsourcing of processes that were traditionally done in-house, particularly among startups and virtual companies with limited lab infrastructure in the small molecule space. Working with external suppliers, he says, allows for immediate access to ready-made and scalable capacity for these types of companies.

To some extent, this trend is borne out by recent developments in the biopharma business space; more than 100 biotech start-ups conducted initial public offerings (IPOs) in 2021(4), almost double that in 2018 (5). While that pace has significantly slowed—just 22 companies went public in 2022, with 2023 on pace for a similar number (6)—it nonetheless indicates that a number of companies have entered the space who may be seeking potential partners for more expansive types of CDMO services, including integrated services.

Eric Langer, president and managing partner at BioPlan Associates, a biopharmaceutical and life sciences consulting firm, noted that data collected by his company reflected an increased reliance in particular forms of outsourcing. In the 2023 edition of BioPlan Associates’ annual 

Biopharmaceutical Manufacturing Capacity and Productio

n market report (7), just 29% of survey respondents indicated that they were doing all of their biomanufacturing for mammalian platforms in-house; this is half the number (58%) of companies who reported similarly in 2006, according to Langer.

“As the complexity of bioprocessing increases, and the need for highly specialized, experienced staff grows in areas like cell therapy, plasmid DNA production, gene therapy, and continuous bioprocessing, and so on, the demand for those unique skills has increased,” says Langer. “Further, many small and mid-sized companies simply do not have the manufacturing core competencies needed to move their clinical projects through the pipeline. For them, there may be no option but to outsource their product manufacturing.”

In addition to the increased types of complex tasks, Langer also noted that the global pandemic shifted the outsourcing models used by some in the market. For example, prior to COVID-19, certain basic operations, like cell culture media and buffer rehydration, were typically an in-house production. Like many other industries, the supply chain shortages caused by the pandemic necessitated consideration of new biomanufacturing channels, including outsourcing.

“The percent of the industry considering outsourced buffer and media services increased dramatically—from single-digit percentages, to a substantial share of the industry now actively outsourcing the production and shipping of buffers and media,” says Langer. “While today this is often done for more complex media and buffers, as the service becomes more common, it is likely to begin replacing some smaller facilities’ media and buffer kitchens.”

Given the various types of expansions occurring in the outsourcing industry, it raises the question of whether potential pharma partners will turn to full-service outsourcing. For companies without existing laboratory or manufacturing services, who are more incentivized to consider outsourcing in some manner, it could make business sense to prioritize a provider who can offer various services. Furthermore, even if clients don’t wish to take advantage of the full “end-to-end” services, using one company for multiple
different tasks will nevertheless offer some level of convenience.

“Demand for companies, including those in the small molecule space, that are able to offer a range of streamlined and efficient integrated capabilities across the development journey is increasing,” says McDermott. “Developers are provided with a single point of contact, which makes it easier to keep track of production timelines. The need for tech transfers is also eliminated, saving time and reducing the risk of error. Silos are reduced and any information or insights gathered in earlier development stages can be quickly shared and actioned downstream, improving efficiency and optimizing the quality of the end product.”

McDermott views integrated CDMOs who can support various development functions, from candidate selection through early development and toward commercialization, as a crucial development for the future of the outsourcing industry. However, there is not a clear consensus on this; even if the industry is facilitating full-service models because of increasingly numerous outsourcing niches, that doesn’t necessarily mean there are clients lining up to take these companies up on said offer.

For instance, according to Langer, the convenience of streamlining development with a single partner needs to be balanced with getting the best value for one’s money in said services. “Sales staff and service suppliers can benefit from the one-stop-shop concepts,” he says. “This concept has been around for many decades, and while many end-users would like to have to deal with just one vendor, it is often unlikely that one vendor can provide the best service at the best price for every one of their services, from cell line development through fill/finish. Typically, a compromise will be seen between the convenience of dealing with one vendor, and the quality, speed, and price of dealing with multiple service providers.”

This type of consideration gets to the heart of what companies are looking for in potential outsourcing partners. In BioPlan Associates’ annual report, 64.2% of the industry valued “supplier’s consistently high products” as a “Most Important” attribute; 50.9% of respondents indicated the same for “performance on vendor audits” (7). In this vein, while convenience matters, it is inferior to broader cost-benefit calculations; a company being local, for instance, typically ranks low on the annual survey for those reasons.

McDermott similarly acknowledged that a company’s reputation was the most important consideration in standing out from the pack. “The main differentiator is whether the company is well established and reliable,” he says. “There are a lot of risks associated with drug development, so forming a partnership with a company that has the experience, specialist expertise, and scope to deliver what they promise helps to mitigate some of the potential issues.”

That said, while companies may not be racing to use the totality of end-to-end services, McDermott notes that they do present an opportunity for outsourcing companies to get their foot in the door. In this regard, offering more services naturally widens the amount of companies who may consider one’s services. In doing so, however, a company must not risk damaging the efficiency of any given service, ultimately inducing a balancing act between offering a breadth of services that attract customers and a quality of service that retains them.

Many find it is difficult to firmly predict where the outsourcing industry will head in terms of integrated CDMOs, given the inherent variability of the pharmaceutical industry. Langer, on the one hand, noted that many CDMOs have adopted a “wait-and-see” approach that “balances the need for establishing expertise in this area against the future developments and demand for such services.”

McDermott, on the other hand, did have pointed predictions, firmly embracing the holistic CDMO model. He characterized traditional outsourcing models, which relied on multiple partners specializing in certain aspects of the development journey, as failing to integrate across drug development functions like drug substance manufacture, clinical trial manufacturing, and clinical testing. Integrated CDMO services, conversely, would provide “centralized program management and control of critical path steps such as the transition into first-in-human or proof of concept studies within a single supplier.”

“Given these benefits,” McDermott added. “It is expected that increasing numbers of companies will look to harness the efficiencies offered by integrated CDMO partners in the coming years.”

While there may be no consensus answer on the future of outsourcing, whether other prominent trends continue in the future may shift the tide one way or the other. Notably, while there is an existing surplus of young biotech start-ups, if venture capital dries up and IPOs continue to dramatically slow-down, the proportion of companies acutely reliant on outsourcing will decrease; this will weaken the prospects of integrated outsourcing companies who can pitch themselves as full-service partners to companies without facilities. If, however, the current state is a valley, the uptick in new start-ups will likely present a boon for companies adopting an integrated model.

Unfortunately for those looking for consensus, experts seem to be divided on the issue, a natural extension of market uncertainties. When discussing a finding from the 2023 Convention on Pharmaceutical Ingredients (CPHI) annual market report concerning biotech funding, Tara Dougal, content director—pharma, Informa Markets, noted that “the next 18 months to two years look extremely robust, and that’s with macro and inflation headwinds factored-in” (8). Whereas, John Stanford, executive director of the Incubate Coalition, a lobbying group focused on the pharmaceutical industry, said in an interview with 

 that adjustments to patent exclusivities from the Inflation Reduction Act added further risk and barriers to entry for venture capitalists looking to invest in biopharma (9).

In this vein, it is pivotal to pay close attention to the market movements in the near future, especially because the outsourcing industry is so heavily predicated on the larger pharma market. While the sustained impacts may not seem evident on the surface, the downstream impact of broader pharma market movements will undoubtedly extend to the outsourcing industry.

1. National Institutes of Health. Decades in the Making: mRNA COVID-19 Vaccines. 

2. Billingsly, A. More Than COVID-19: 6 Other Promising mRNA Vaccines in the Pipeline. 

3. Vickers, J. et al., Overcoming Engineering Challenges to Enable Commercial Scale mRNA Vaccine Manufacturing. 

4. Brad Loncar. Biotech IPO’s Class of 2018. 

5. BDO USA. BDO Biotech Brief – Q1 2023. 

6. BioPharm Catalyst. Biotech IPO Calendar. 

Report and Survey of Biopharmaceutical Manufacturing Capacity and Production

. 20th edition; BioPlan Associates, Spring 2023.

8. Playter, G. Predicting Pharma’s Future. 

9. Playter, G. The United States and the Global BioPharma Market. 

When referring to this article, please cite it as Playter, G. Investigating Outsourcing. 

Articles

Cover Story

Broader industry shifts may lead outsourcing companies to prioritize integrated services.

Investigating Outsourcing

Review articles are quick to outline the tremendous progress and consolidated gains accrued during the past 40 years for cell and gene therapies (CGT). Because they are retrospectively oriented, review articles are not always adept at identifying means to further optimize performance. For example, of the approximately 1000 known adeno-associated virus (AAV) variants, “…our knowledge of AAV’s infection pathway is based on only a few serotypes. The prevailing understanding is that serotypes bind to different, or have differential affinities to an array of primary cell surface glycoprotein receptors and secondary receptors” (1).

Pharmaceutical research energy is now being focused on studies to elucidate if the lower performance of therapies for some racial minorities or gender are based on these serotype differences (2).By studying these less understood serotypes, it may be possible to uncover a mechanism that not only works for more patients, but might just work better per se. This could be an opportunity similar to gene editing with Cas9 proteins, which was admittedly partly driven by a desire to find paths around patent claims. Today, NgAgo protein gene editing systems (argonaute family proteins) are able to cut DNA at a specific site without requiring guide RNA. And relevant to CGT, mini-Cas9 are derived from staphylococcus aureus bacterium, which are small enough to fit inside these viruses, alongside the increasingly popular Cpf1 (clustered regularly interspaced short palindromic repeats [CRISPR] from 

When immersed in a topic-specific scientific paper (not a review paper), it quickly becomes clear how enormous a difference AAV2 binding to heparan sulfate proteoglycans is, compared with, say, binding to angiotensin-converting enzyme 2 (ACE2) and the SARS-CoV-2 spike protein, which enhances COVID-19 virulence (2). Indeed, the spike protein for COVID-19 was initially rumored to have been a lab-engineered version of the HIV spike protein, atypical of coronavirus, and not a naturally occurring variant.

Harnessing a virus to alter a patient’s genetic code as a therapy will always present natural obstacles cojoined to natural fear. CGT has a deep foundational knowledge accrued during the study of HIV and similar pandemics. Harnessing these viruses brings along some of the problems they create for a human’s complex immune system responses. Most people carry neutralizing antibodies (NAbs) against one or more of the commonly used vectors and capsids, thus greatly reducing efficacy for those individuals.

According to a study, racial minority and gender differences were a determining factor in efficacy. The study concluded, “We suggest that capsid selection for drug development take into consideration AAV seroprevalence among the patient population while avoiding sampling bias” (4). Understanding the underlying biology in subpopulations may lead to more highly optimized treatments.

Naiara Tejados, head of Communi-cations, VIVEBiotech, says, “Regarding adverse immune responses, compared to other vector types, only a small portion of the population has preexisting antibodies to lentiviral vectors [LLVs]. As a result, patients who receive CGTs based on lentiviral vectors will have low risk of experiencing immediate immune reactions.”

Hanna Dreja, Upstream Subject Matter Expert at Astrea Bioseparations, summarizes her perspective on balancing CGT use applications saying, “Lentiviral vectors (LVV) are widely used in cell therapy, specifically for autologous treatments, where a patient’s own cells are infected with viral particles and then reintroduced into the body for chimeric antigen receptor T Cells (CAR-T) and stem cell therapy. Conversely, AAV vectors are directly administered to patients to deliver therapeutic genes.” When it comes to examining performance against immune response, Dreja goes on to say, “While the immune response to LVV is generally less concerning, repeated AAV injections can lead to side effects and reduced therapy effectiveness, due to the presence of neutralizing antibodies. However, it is important to note that LVV carries the added risk of integrational mutagenesis. Despite these concerns both systems are good solutions for long-term and stable expressions of the desired genes.”

Manufacturing CGTs at a large scale poses significant challenges. Automation and improved process analytics that provide data-driven decision making leads to improved efficiency and cost reductions. According to Tejados, “the main advantage of allogenic gene-modified cell therapy products is the decrease in manufacturing costs. Manufacturers in the field of gene and cell therapy are making great efforts to lower the costs of advanced therapies, and obviously allogeneic therapies are more accessible,” she says. Tejados also points to some customer projects using intermediate-scale bioreactors that have a small footprint that reduces costs at development stages.

Dreja states that, “Recent advancements in single-use units and automation have made it possible to reproducibly generate these particles consistently, although at a high cost. Inducible packaging cell lines offer an intriguing alternative to transient transfection protocols, as they eliminate the need for large quantities of expensive plasmid DNA.” Tejados points to recent successes with stable producer cell lines, where there is obvious industry excitement mixed with some hesitation.

Luca Peterfay, process development scientist II at Charles River, however, emphasized that the generation of the stable producer cells is an additional step prior to the initiation of production, “…and this step can potentially increase preclinical timelines by several months. The generation of stable producer cells is a complex and time-consuming procedure, which entails tedious cell-line development due to the numerous clonal isolation steps followed by extensive screening procedures,” she explained (5).

For the constitutive approach, Peterfay also noted, “… due to replacement of the VSV-G with a low-cytotoxicity alternative envelope, titers may not be as high as those achieved via transient transfection, and transduction efficiency can also be lower. The limitation in titers can be compensated by longer processing times and multiple harvests; however, due to the reduced transduction efficiency higher doses might be required to achieve the required therapeutic effect in patients,” she said (5).

Developing a purification system that is effective and simple is crucial as LVVs in particular are delicate molecules and suffer under processing sheer forces and stresses and, therefore, benefit from minimal handling. “Even more effective and gentle approaches, which eliminate contaminants, will be key for future production at scale,” Dreja suggests. “For example, next-generation purification technologies, such as nanofibers, present an interesting solution for LVV and AAV purifications, offering an open structure and a large surface area that is easily accessible to viral vectors. The nanofiber technology has the potential to reduce processing times by relying on convection-based mass transfer rather than diffusion,” she concludes.

1. Bulcha, J.; Wang, Y.; Ma, H.; Tai, P.; and Gao, G. Viral Vector Platforms within the Gene Therapy Landscape. 

. 2021, February Springer Nature Group.
2. Mengstie, M. Viral Vectors for the in Vivo Delivery of CRISPR Components: Advances and Challenges. 

Frontiers in Bioengineering and Biotechnology

, May 2022 Frontiers.
3. Clausen, T.; Sandoval, D; Spliid, C. et al. SARS-CoV-2 Infection Depends on Cellular Heparan Sulfate and ACE2. 

. 2020, September 12;183(4):1043-1057.e15. Elsevier DOI: 10.1016/j.cell.2020.09.033
4. Khatri, A.; Shelke, R.; Guan, A.; Somanathan, S. Higher Seroprevalence of Anti-Adeno-Associated Viral Vector Neutralizing Antibodies Among Racial Minorities in the United States, 

. 2022 April 33(7-8). Doi: 10.1089/hum.2021.243
5. 

, C. State of Producer Cell Development for Lentiviral Vector Manufacturing. 

Chris Spivey is the editorial director of 


Vol. 36, No. 8
August 2023
Pages: 13–14 

When referring to this article, please cite it as Spivey, C. Balancing AAVs Versus LLVs. 

Development

Burgeoning CGT modalities bring promise and challenges.

Balancing AAVs Versus LLVs 

Optimizing cell-culture processes for monoclonal antibody (mAb) production is a key factor in scaling up manufacturing to commercial levels. To maximize cell-culture output, strategy is required that takes into consideration factors such as cell culture media, environmental monitoring, and process controls. Yet, despite decades of process optimization, there remain stubborn challenges—or new challenges that have arisen–during the scale up of cell culture processes.

Previous process optimization in mAb production meant that mAb titer and cell density were once low, and a successful scale up was determined through quantity, that is, achieving the same mAb titer at larger scales, notes Masa Nakamura, PhD, global bioprocess science specialist at 3M Separation and Purification Sciences Division. However, he continues, now that the biopharma industry is in an era of higher titer and cell density, the focus has shifted from “quantity” to “quality.”

“Engineered modern mAb products require higher degrees of process control to produce high-quality products along the scale up, which demands a holistic approach in cell-culture scale up,” Nakamura says.

Not only do biomanufacturers need to control cell densities and titers, but they must also manage post-translational modifications such as glycosylation profiles, says Susan Riley, leader, Cell Culture business, Cytiva. As titers have increased, the balance of the bioproduction workflow must also be well managed, she states.

“It does little good to increase the expression of protein to 10 g/L [for example] if significant yield will get lost in the downstream filtration, purification, or even final filling steps,” Riley cautions. In terms of upstream hardware—with the possibility of failure, even in late-stage clinical trials—Riley notes that biomanufacturers want to minimize risk by delaying critical capacity decisions until as close to final approval as possible.

Andy Topping, PhD, chief scientific officer at FUJIFILM Diosynth Biotechnologies, points out that, over time, as expression systems have become more efficient, the viable bioreactor scale has decreased, with many commercial products able to be produced in few 2000-L single-use bioreactor runs per year and the industry has responded by developing both scale-up and now scale-out strategies. This trend has placed enormous pressure on solution management and downstream processing in legacy assets, and novel fluid management systems, including in-line dilution and blending, are now maturing. “As a result, scale and capability of single-use process systems with full process closure are continuously expanding,” says Topping.

Meanwhile, processes that are expected to require manufacture at very large scale will often need to be transitioned from single-use bioreactors to stainless-steel bioreactors, Topping emphasizes. Although recent innovations have made the performance of these single-use systems closer to stainless steel, many processes will still have been developed and operated in bioreactors with fundamentally different engineering, mixing, and gas transfer capabilities, where harvesting will be conducted by depth filtration rather than centrifugation. Scale up of these processes therefore also means translating the process correctly to the new systems, according to Topping.

“Moving forward, the industry finds itself in a secular trend of increased diversity as more capable systems and fundamental understanding become available at an accelerated pace,” says Thomas Page, PhD, vice-president of Engineering and Development, FUJIFILM Diosynth Biotechnologies.

Multiple innovations have arisen for the sake of making cell culture scale-up more efficient. According to Topping, the most fundamental innovations have been improved cell lines and media, which enable the higher cell densities and higher titers. That increase, in turn, allows many processes to use a scale-out approach at or near to the scale used for clinical supply, rather than a scale-up approach.

Single-use manufacturing systems are also an innovation that by design enables an optimum solution for closed processing and better aseptic techniques as well as ease of use. Single-use platforms offer both standard and flexible workflows. Furthermore, the impact of having a closed process reduces contamination risk while enhancing efficiency and predictability, benefits that cannot be overstated, says Page.

“For example, use of full isolators for flask work significantly decreases flask contamination rates and resultant schedule and manufacturing cadence impacts. Integration of standard and custom sensors with the single-use consumables further reduces risks of contamination,” Page states.

“Full understanding through detailed risk assessment of process closure allows facilities to operate with multi-product cell culture on a predictable high cadence rate. This significantly reduces overhead and significantly increases availability of capacity to the market,” Page adds.

Meanwhile, innovations for bioreactors include new fluid mixing formats that create more efficient oxygen transfer with lower shear; this helps to support higher oxygen demand for both intensified and perfusion cultures, says Nakamura. Outside of the bioreactor and when looking at the harvest and clarification of cell cultures, single-use centrifuge and chromatographic clarification are new additions to the bioprocess toolbox for high density cell culture scale up, he adds.

“However, for cell retention in perfusion cultures, the cell retention devices would need to evolve further to include more precise pore membrane structures that achieve high precision separation with minimal clogging,” Nakamura states.

Page makes the observation that growth in the cell therapy space, which has resulted in the concomitant development of robust cold storage containers for mammalian cells, has opened the door for the use of high-density cell banks. “High-density cell banks offer the potential to massively decrease labor, risk, and time in plant. It is a critical capability for maturing clinical phase perfusion offerings,” Page states.

Other developments, such as novel sensors (e.g., Raman soft sensors), have allowed greater real-time insight into process performance at all scales, Topping also states. These novel sensors are being applied in the continuous monitoring of metabolites, such as glutamine and lactate, as well as mAb titers in the bioreactor, Topping explains.

“Other emerging technologies benefiting large-scale biotherapeutic manufacturing are artificial intelligence data science for integrated processing modeling, robotics, automation, and the related transition to paperless program management and data management systems,” says Page. Digital twins, for instance, offer the potential for enhancing both speed-to-market and post-approval optimization. In-silico modeling offers the potential to select optimal cell lines and modified sequences that can optimize across a broad arc—from potency to selectivity to manufacturability. All these benefits offer the promise of increased patient access and sustainability, adds Page.

Innovation compatibility with single-use technology

Vaccines were traditionally manufactured in chicken eggs or in stainless-steel systems, but the expansion into single-use technologies, and particularly the growing adoption of single-use bioreactors, has helped transform the global capacity for vaccines, both in terms of efficiency and flexibility, Riley points out.

Specifically, Cytiva has expanded the single-use technology approach to include viral vector-based therapeutics, including oncolytic vaccines, in the design of its prefabricated, modular facility (KUBio, Cytiva); recent expansion of its modular facility design is aimed at accelerating the manufacture of and reducing the risks of viral vector-based therapeutics.

Nakamura, meanwhile, notes that innovations in the harvest and clarification of cell cultures involve single-use technologies, specifically single-use centrifuge and chromatographic clarification processes. “These technologies are compatible with current single-use bioprocess workflows and would often provide process economic advantages for mAb production in high density cell cultures,” he states.

Page emphasizes that it is now possible to engineer large, capable single-use assets that are fully electric and devoid of any steam production. This innovation, coupled with high density production and process closure means that energy usage could be decreased up to an order of magnitude; however, such an energy decrease will need to be verified as next-generation plants come online, Page explains.

It is important to keep in mind that many innovations taking place are agnostic to the production platform, but some are specific to either a single-use or stainless-steel system. Improved bioreactor performance, increases in scale, development of more single-use harvesting options, and novel cell retention devices for perfusion have all pushed bioproduction capabilities forward, according to Page.

“In general, it is important for any company working in this space to have a good risk-based model to understand the type, scale, and pathway for hazards and to have a clear, explicit control strategy. Aligning this understanding end-to-end in an organization is critical,” says Page.

For example, to optimally move a development project from clinical single-use manufacturing through to large-scale stainless-steel manufacturing, one needs to understand the impacts of product and unit operations. This understanding must then be mapped out to differentiate between product-specific requirements and facility and equipment controls. This mapping must be done in such a way that the chemistry, manufacturing, and controls sections may be developed to fit the target assets used for the product lifecycle, explains Page.

“With the target lifecycle map or potential pathways mapped, process knowledge may be subjected to directed evolution such that costly remediations, such as equipment changes, validation extensions, or redevelopment may be avoided,” says Page.

Another area for mapping back and forth between single-use and stainless-steel systems requires the development of unit operations that were previously unavailable in a single-use system. These include continuous centrifugation and homogenization, for example, says Page. However, these topics go beyond mAb processing and will have profound impact on advanced therapies, microbial products, and vaccines.

“The key is for industry to have a mappable process that allows easy transition into and then later back out of large-scale stainless-steel production and product lifecycle. Ideally each product could be produced in its most efficient embodiment over this lifecycle. As these capabilities mature, there will clearly be opportunities for hybrid deployment,” Page states.

To have successful scale-up, it is imperative that product consistency, in both quality and quantity, be addressed. Having product consistency requires a holistic approach in the upstream stage, the clarification step, and downstream purification, notes Nakamura. Focusing solely on mAb titer would not be enough to provide successful scale up and consistent manufacturing.

“An example of an effective strategy in scaling up mAb cell culture to commercial scale includes optimizing cell culture from both upstream and downstream point of views,” Nakamura intones. “Cell culture is not only the source of products, but also the source of all soluble and insoluble contaminants (cells, debris, host cell proteins, DNA, product variants), which need to be removed in downstream. Harvest criteria and timing can be intentionally selected to balance the high titer and low contaminants to achieve the most overall benefits.”

In Cytiva’s experience, Riley notes, the most effective strategies for successful cell culture scale-up involve a deep understanding of how the entire bioproduction workflow must work together in a balanced way. “Paying attention to the vortex speed or air sparging in the bioreactor, the crossflow and pressures across TFF [tangential flow filtration] membranes, and the aspect ratios associated with resins in purification columns all contribute to the ultimate yield of the final product. And at the end of the process, if the therapeutic is of absolute premium value, such as in gene therapy, the use of robotic aseptic filling [is important] so that not a drop will be lost,” she explains.

Riley further notes that automation and digital strategies are solutions that can be deployed to streamline the cell-culture process, enable the continuous manufacturing concept, reduce process variability, and lower production costs. “It should be noted that collaboration between tool/solution providers and therapeutic developing organizations is greatly beneficial for accelerating the technology advancement,” she adds.

As molecules diversify and precision medicine continues to advance, says Riley, there will be smaller production runs that require maximum flexibility, and a variety of single-use components will become increasingly important. “And, frankly, some of the newer modalities use little cell-culture media at all but require a deep understanding of the role that other fluid components can play in processing,” she states.

Meanwhile, Page explains that traditional strategies, including mapping exercises from scale-to-scale, allow small-scale design of experiments, which are representative of production-scale parameter space and therefore useful. In the past, stainless-steel bioreactors were designed within a relatively narrow range of capability, but today, the advent of single-use bioreactors has required the industry to develop ways of migrating from stainless-steel to single-use systems as well as across different vendor-supplied single-use bioreactors.

“Today, many programs may migrate across different bioreactors over their lifecycle. The design-of-experiment approach will continue and will likely be enhanced as next-generation single-use bioreactors with enormous operation space become available,” says Page.

The key issue for contract development and manufacturing organization (CDMO)-specific operations today, as Page sees it, is for CDMOs to select equipment that have the broadest operating space and to nest such equipment in a flexible facility. Facilities must increasingly have the broad capability to efficiently support different modes of operation, Page concludes.

Feliza Mirasol is the science editor for 


Volume 36, No. 8
August 2023
Pages: 15–18

When referring to this article, please cite it as Mirasol, F. A Look into Biologic Scale-Up Strategies. 

Upstream Processing

Scale-up strategies for cell culture will require a deep understanding of overall bioprocess workflows.

A Look into Biologic Scale-Up Strategies

As biopharmaceutical manufacturers seek greater efficiency and intensification of downstream processes, process automation stands ready to offer improvements. Automated process control enables efficient operation of connected systems and reduces variability and error. Process analytics collect and analyze large amounts of real-time data to improve process monitoring and control. Process automation, as well as robotic handling in some cases, facilitates the use of closed systems that reduce the risk of manual intervention. Smaller footprints and faster production boost capacity to meet the increasing need for biopharmaceutical therapies.

The Sartorius Resolute BioSC platform intensifies downstream processing using an automation system to connect steps including chromatography, virus inactivation, and polishing. The connected system reduces the equipment footprint and process time. For example, Sartorius collaborated with Enzene Biosciences to improve a monoclonal antibody (mAb) purification process (1). First, a batch process was developed to remove the intermediate buffer adjustment step. Next, the unit operations were run in a connected mode, using a fully automated BioSC modular system, which reduced the overall equipment footprint by approximately 80% and processing time by approximately 40%, the company reports in an interview with BioPharm International.

In an ongoing collaboration between Rooster Bio and Sartorius, the companies are working together to commercialize a human mesenchymal stem/stromal cell (hMSC)-based exosome production platform (2). Sartorius’ automated, batch benchtop filtration systems (Sartoflow Smart and Sartoflow Advanced) will be used in the downstream processing operations. These tangential flow filtration (TFF) systems are optimized for ultrafiltration and diafiltration applications and already used in many downstream processes, such as purification of vaccines, mAbs, and recombinant proteins.

Cytiva’s automated, single-use filtration system (ÄKTA readyflux) is used in TFF operations and in normal flow filtration. One or more TFF units are used in workflows to manufacture many types of biopharmaceuticals, including mAbs, plasmid DNA (pDNA), viral vectors, and mRNA. The equipment is available with different industrial automation control solutions. “Unicorn is the standard control, but PlantPAx via Unicorn OPC [open platform communications] connection and DeltaV automation systems are also available. The system can be a stand-alone unit operation, connected via OPC to a historian for data integration, or [have] full integration via Unicorn OPC into a [distributed control system] layer,” reports the company in an interview with BioPharm International. “Automation, across the workflow, can improve quality by reducing variability and errors.” In February 2023, Cytiva announced that Australian contract development and manufacturing organization AcuraBio would use Cytiva’s two-step purification protocol and ÄKTA readyflux in a FlexFactory configurable manufacturing train to manufacture pDNA (3).

In downstream processing of biopharmaceuticals, such as mAbs, there is a continuing push to greater efficiency through process intensification using single-use, multi-product facilities as well as more data analytics and process automation. For single-use systems, it is crucial for systems to be handled and integrated properly. Proper installation and use are facilitated by appropriate physical design and other features, such as physical and electronic tags on the system and the assembly.

Stuart Tindal, chair of the Bio-Process Systems Alliance automation committee and product manager of Intensified Downstream Systems at Sartorius, adds that although installation and tear-down of single-use systems is primarily manual, process automation systems can now support operators performing these functions. For example, step-wise user guidance can be communicated from the human-machine interface, and tools such as contact verification sensors can be used to ensure that connections are made and are secure. Through digital linking and single-use consumable design, the automation system can identify the single-use assembly and upload the material article and lot number into the system batch details via a camera and label-text and code recognition, explains Tindal. Automation thus reduces the risk of transfer error that is inherent in a human reading and entering information.

Darren Verlenden, head of Process Solutions at MilliporeSigma, adds that because an important advantage of single-use bioprocessing is flexibility, the automated system must be designed to easily adapt to different assembly configurations, bag volumes, and tubing sizes. In addition, the sensors that are part of automated processes (e.g., non-contact flow meters, pressure probes, pH probes) must be compatible with single-use flow paths.

Process automation requires a shift away from in-process manual sampling with off-line analytics toward in-line or on-line process analytic technologies (PAT) that enable continuous measurements of critical parameters and critical quality attributes (CQAs), says Verlenden. Spectroscopy continues to be an important in-line measurement tool for monitoring CQAs in bioprocessing. “We are seeing continued advancement of ultraviolet, infrared (IR), near-IR, and Raman spectroscopies. In particular, Raman spectroscopy holds the potential for providing the most complete molecule and impurity information, while being minimally impacted by water signal. [MilliporeSigma’s] ProCellics Raman analyzer can provide continuous and immediate insights into protein concentration, aggregation, and the presence of impurities.”

In cases where in-line technologies are not compatible with analytical requirements, MilliporeSigma’s Modular Automated Sampling Technology (MAST) allows samples to be extracted and delivered to analytical instruments for “near real-time analysis” while maintaining sterility, adds Verlenden. Automated, sterile sampling is gaining traction due to the growing interest in closed processing, he explains.

“As more novel modality therapeutics are moving to the manufacturing floor, there is strong interest to deploy and adapt the PAT tools to them,” suggests Verlenden. “Technologies like automated sampling are modality-agnostic and could be readily used in novel bioprocesses such as viral vector manufacturing.”

MilliporeSigma and Agilent announced a collaboration in June 2022 (4) that included work to demonstrate the utility of on-line high-performance liquid chromatography (HPLC) to monitor and control antibody aggregation levels in bioprocessing. Because aggregation can impact biological activity of the biopharmaceutical, ensuring that aggregates are sufficiently reduced during the process is crucial for ensuring product quality, explains Verlenden. The companies used Agilent’s on-line HPLC as PAT to monitor antibody aggregation levels after flow-through frontal chromatography purification using a cation exchange resin.

“Upon detection of aggregates in the column effluent, automated column switching was initiated to ensure effective and efficient continuous removal of aggregates from the process stream. This study provided a solid proof of concept of physical and digital integration of a processing technology (cation-exchange chromatography for aggregate removal) and on-line analytical technology (HPLC for aggregate detection), which now could be extended to more complex CQAs. We look forward to continuing this collaboration to expand the use of on-line HPLC along the broader bioprocessing workflow and in conjunction with other analytical

This broader implementation of PAT will enable the industry to make progress toward enhanced process monitoring and control of downstream processes. “The implementation of PAT enables continuous manufacturing and empowers industry progress towards Bioprocessing 4.0,” Verlenden concludes.

Automation is crucial for cell and gene therapy (CGT) manufacturing, where demand currently outstrips supply; closed, end-to-end robotic automation platforms aim to create a step change in capacity (5). Automated and closed unit operations make a difference in both integrated operations and in separate upstream and downstream operations.

Thermo Fisher Scientific introduced the Gibco CTS DynaCellect Magnetic Separation System (DynaCellect), an automated and closed system for cell isolation, cell activation, cell depletion and Dynabeads magnetic beads removal, in October 2022 (6). The new system eliminates the manual intervention required with the earlier CTS DynaMag tool. “DynaCellect is designed to be an automated accessory for workflows that leverage DynaBeads,” says Evan Zynda, senior staff scientist at Thermo Fisher.

The new, scalable system can help cell therapy developers move from process and clinical development to commercial manufacturing. It is currently used in T-cell therapy manufacturing workflows and is being used in developmental workflows for many other therapy types, says Zynda. The isolation protocol fits a broad target cell range and can be used in autologous and allogeneic processes.

“Scalability stands out as a major consideration when developing a product that can accommodate autologous and allogeneic processes,” says Zynda. “As the industry continues to progress, autologous groups desire to scale down the manufacturing volume and time of these personalized workflows, [which] will allow for a more rapid turnaround time, reduced costs, and easier downstream processing. On the other side, allogeneic processes will be longer and scaled up to create larger batch sizes that can result in several banked doses.” Zynda explains that downstream processing for these larger batch sizes, anticipated for products such as natural killer-cell therapies and induced pluripotent stem cell-derived therapies, could be more cumbersome. He says, however, that the tool will be designed to meet this growing need. “At launch, the DynaCellect could easily accommodate a range of 100 million to 10 billion target cells for isolation, which is compatible with almost all current isolation processes. We have since validated targeting 20 billion cells of interest without issue, and we will continue to develop protocols to push the system further as the need arises.”In addition, the continuous de-beading process can accommodate any volume of bead-containing solution, including smaller autologous batches and larger allogeneic batches, says Zynda.

Zynda adds that automated workflows will be beneficial for decentralized or “point-of-care” manufacturing sites, which are less likely to have specialized, trained technicians. “Creating a closed and automated workflow not only enhances efficiencies, it also increases cell therapy manufacturing success rates and reduces failures,” he adds.

Technology provider Invetech also sees a need for modular solutions for unit operations that can speed up the manufacturing process for cell therapies (7). The company’s Korus closed system for elutriation and cell wash using gentle counterflow centrifugation was launched in June 2022 (8). “Manufacturing efficiency of cell therapies needs improvement and innovation if the cell therapy industry is going to reach its commercialization goals,” said Jon Ellis, lead of Invetech’s Cell Therapy Science and Application team, in the press release (8). “Our data shows that whilst achieving similar apheresis lymphocyte cell recovery to the control wash process, Korus eluted lymphocytes to high purity, which resulted in improvements in downstream performance including greater Dynabead cell selection recovery and fold expansion.”

Sartorius. What are the Benefits of a Connected Downstream Process using Resolute BioSC? Webinar, March 29, 2023.

Sartorius. Sartorius Collaborates with RoosterBio to Advance Downstream Purification Processes for the Manufacture of Exosomes. Press Release, Jan. 5, 2023.

Cytiva. AcuraBio Expands cGMP Plasmid DNA CDMO Services with Cytiva’s Latest Single-use Purification Technology. Press Release, Feb. 9, 2023.

MilliporeSigma. MilliporeSigma Announces Collaboration with Agilent Technologies to Fill Industry Gap in Process Analytical Technologies for Downstream Processing. Press Release, June 14, 2022.

Markarian J. Automation Aids Cell and Gene Therapy Production. BioPharm International 2023 36 (7).

Thermo Fisher Scientific. Thermo Fisher Scientific Launches Closed and Automated Cell Isolation and Bead Removal Solution to Help Evolve Cell Therapy Manufacturing. Press Release, Oct. 16, 2022.

Invetech. Where to Next in Cell Therapy Manufacturing? Invetechgroup.com, Sept. 19, 2022.

Invetech. New Korus Technology Set to Transform Industry Standards in Elutriation and Cell Wash. Press Release, June 21, 2022.

Jennifer Markarian is manufacturing reporter for BioPharm International.


Volume 36, No.8
August 2023
Pages 19-21 

When referring to this article, please cite it as Markarian, J. Streamlining Downstream Processes. 

Downstream Processing

Process automation technologies aid modernization and enable closed systems.

Streamlining Downstream Processes

Allogenic cell therapies produced in large batches from healthy donor cells have the potential to address many of the logistics and manufacturing challenges associated with autologous cell therapies. They face their own production hurdles, however, including the need to generate large homogeneous cell populations while maintaining functionality and avoiding phenotypic variability. Several strategies are being explored to address these issues while also ensuring the development of cost-effective processes and streamlined development and manufacturing timelines.

All allogeneic cell therapies, regardless of the cell type and whether they are genetically modified or not, face some fundamental challenges. These challenges include sourcing of sufficient, suitable cells, access to fit-for-purpose reagents and manufacturing systems, and scalability of upstream, downstream, fill/finish, and cryopreservation processes.

Unlike autologous therapies, which use cells from the patient being treated, allogeneic therapies are manufactured in large batches from unrelated healthy donor tissues such as bone marrow or lymphocytes, including natural killer (NK) cells and T cells. “This approach makes allogeneic therapies more attractive, but introduces new complexities related to sourcing and handling of cells from different donors​ that can trigger immune responses in the recipient’s body,” notes Nikhil Tyagi, director of cell therapy process development at Center for Breakthrough Medicines. He adds that cells from different donors will have different propensities for expansion and therefore yields.

Cell-therapy developers also must work with technologies for large-scale manufacturing that are not yet fully optimized for the development and manufacturing of allogeneic products, notes Tatiana Nanda, head of process development–cell therapy, drug product and process analytics at Center for Breakthrough Medicines. “Innovations in areas such as cell culture media and manufacturing equipment are required to increase the efficiency, safety, and yield of the process,” she contends. In addition, Nanda believes that optimization of culture conditions, culture media, growth factors, and implementation of automated manufacturing processes are necessary to address this challenge.

Scaling up cell production, particularly for adherent cells, presents another challenge. “Current expansion methods often rely on labor-intensive and manual processes that are not easily scalable. Developing robust and efficient manufacturing processes that can produce sufficient quantities of cells is a critical requirement in today’s world,” Tyagi says.

Even in cases where large-scale allo cell expansion is achievable, there are still limitations with respect to efficient harvest of high numbers of cells, according to Nanda. She also notes the need for ultra-fast fill/finish, labeling, and visual inspection processes which frequently are limited to a 30–180-minute time window. “High-throughput fill/finish and automated visual inspection capabilities can resolve these bottlenecks,” she says. Custom solutions for large-scale, robotics-enabled cryopreservation also help with cryopreserving large batches of allo drug products.

The quality and composition/variability of the donated cellular starting material are of paramount importance in allogeneic cell therapy. “These factors can significantly impact the success or failure of large-scale cell expansion,” states Tyagi.

The quality of donated cells directly impacts the safety and efficacy of allogeneic cell therapies, as poor-quality cells may not function as intended, leading to suboptimal therapeutic outcomes. In addition, Tyagi observes that improperly screened cells could potentially transmit infectious diseases.

The composition of the cellular material, meanwhile, can significantly influence the therapeutic effect, because different cell types have different roles and functions. “For instance,” Tyagi says, “the presence or absence of certain cell types, such as T cells in hematopoietic stem cell transplantation, can contribute to complications or beneficial effects.” Inter- and intra-donor variability, meanwhile, can affect the consistency of allogenic cell therapies.

The methods used to process cells (isolate, purify, and store) after donation can also affect their quality and function, including their viability and therapeutic potential, according to Tyagi. He stresses that standardized, validated cell-processing methods are essential to ensure the consistency and effectiveness of allogeneic cell therapies.

Finally, Tyagi notes that the impact of the quality and composition/variability of the starting material can vary depending on the cell type, the need for genetic modification, and the ultimate application of the cell therapy. “Certain cell types may be more sensitive to the quality of the starting material or more prone to variability. Similarly, the need for genetic modification can introduce additional complexity and potential variability,” he explains.

Finally, the ultimate application of the cell therapy can also influence the importance of the quality and composition/variability of the starting material, according to Tyagi. For example, therapies intended for life-threatening conditions may have stricter requirements for the quality and consistency of the starting material.

Ensuring homogenous expanded cell populations

Several strategies are used to ensure that expanded cell populations have an appropriate level of homogeneity. Given the variability in donor cells used as starting materials for cell therapy, cell sorting is widely used because it is an essential technique for isolating specific cell types from a heterogeneous population, according to Tyagi. “By relying on specific cell-surface markers, these techniques, such as fluorescence-activated and magnetic-activated cell sorting (FACS and MACS, respectively), allow for the identification and separation of desired cell populations, ensuring the production of homogeneous cell populations for downstream applications,” he says.

Genetic modification is another approach that enables the engineering of cells to express specific markers or proteins. This technique, Tyagi observes, facilitates the identification and isolation of target cell types, enhancing the homogeneity of the population. Genetic modifications can also improve cell functions and properties for specific applications, further optimizing their suitability for desired therapeutic or research purposes, he adds.

Manufacturing approaches impact the homogeneity of expanded cell populations as well. The composition of the culture medium, the choice of substrate for cell attachment in adherent processes, and physical process parameters during cell culture, such as temperature and dissolved oxygen content, can influence cell behavior and differentiation, according to Tyagi. “By carefully controlling these factors, researchers can create an optimal environment that promotes the growth and differentiation of desired cell types, which helps to maintain the homogeneity of the cell population and ensures consistent and reproducible results,” he contends.

Use of microfluidic devices, meanwhile, provides a means for gaining precise control over cell isolation and culture conditions, Tyagi comments. They can, for instance, isolate specific cell types based on physical properties such as size and deformability. “By culturing cells within microscale environments, these devices provide tight control over culture conditions, enhancing the homogeneity of the cell population,” he says.

It is also worth noting that different combinations of techniques are used to ensure homogeneity after expansion of specific types of cells. For mesenchymal stem cells (MSCs), according to Tyagi, culture conditions play a significant role in maintaining stemness and preventing differentiation. For T cells, genetic modification combined with cell sorting techniques has been a key strategy, particularly for chimeric antigen receptor (CAR) T-cell therapies. Cell sorting is also crucial for isolating hematopoietic stem cells (HSCs). For dendritic cells, a combination of culture condition manipulation and genetic modification is often employed, while for natural killer cells, all three approaches (cell sorting, control of culture conditions, genetic modification) contribute to more homogeneous expanded cell populations.

Maintaining cellular integrity and functionality

Expanded cell populations must not only be homogeneous, but have a high percentage of viable cells with the desired functionality. Many different factors can influence the ability to maintain these attributes during expansion. First, says Tyagi, is the cell source and type. For instance, pluripotent stem cells (PSCs) require a different set of culture conditions compared to somatic cells like T cells or MSCs. “PSCs need a feeder layer or feeder-free conditions with specific growth factors, while T cells and MSCs can be expanded using media supplemented with serum or serum-free conditions,” he explains.

For genetically modified cells, optimization of the mechanism for delivery of the genetic material (e.g., viral vector, electroporation, etc.) is essential, as these methods can impact both cell viability and functionality, according to Nanda. As an example, she points to CAR T-cell therapy production, which involves T-cell activation, genetic modification to express the CAR, and then expansion of the CAR-T cells.

Not surprisingly, the choice of expansion system can also affect cell integrity and functionality. Traditional planar systems, Nanda notes, have limitations in terms of scalability, so suspension culture systems and bioreactors are being explored for large-scale production. She adds that these systems also allow for better control of culture conditions.

As with any drug manufacturing process, quality control (QC) throughout the cell expansion process is a must. “QC measures, including monitoring of cell identity, purity, potency, and safety, are essential to ensure that cells intended for use as cell therapies maintain their desired characteristics,” Nanda says. Advanced techniques like flow cytometry, next-generation sequencing, and functional assays are used for these purposes.

Finally, the use of automation helps to reduce variability and increase reproducibility, while closed systems minimize the risk of contamination. After expansion, appropriate cryopreservation and thawing processes must be established and carefully managed to maintain both cell viability and functionality.

During expansion, it is possible for some cells to undergo changes in their phenotype. For cell therapies, it is essential that all cells in the expanded population have the desired phenotypes. It is therefore necessary to take steps to avoid differentiation. There are several strategies employed to achieve this goal, according to Tyagi, and the specific strategy employed can depend on the cell type, the need for genetic modification, and the ultimate application of the cells.

Optimized culture conditions with cell-specific culture media, growth factors, and cytokines can help maintain the desired cell phenotype during longer expansion periods, Tyagi says. Use of bioreactors, for instance, allows better control of process parameters such as oxygen levels, pH, and nutrient supply, which can influence cell differentiation. Hypoxic conditions, meanwhile, can activate certain signaling pathways that promote stemness and inhibit differentiation. Cells can also be genetically modified to alter specific genes that control cell differentiation to increase the stability of the desired phenotype. Co-culturing stem cells with other types of cells can help maintain their stemness, while 3D culture systems provide a more in vivo-like environment for cells, including favorable cell-cell and cell-matrix interactions.

Maintaining aseptic conditions during longer cell expansion processes poses challenges, but several strategies are being employed to address them. Top of the list, according to Nanda, are the use of automated and closed systems with stringent quality control measures in place. Operator training, facility design, and the use of disposable production systems are also important.

Automated systems, Nanda says, control and monitor culture conditions in real-time, allowing for adjustments in parameters such as temperature, pH, and nutrient supply to ensure optimal growth conditions for the cells. They also reduce manual handling, minimizing the risk of contamination. Closed systems also reduce the potential for contamination by isolating the the cell culture from the external environment, as do stringent quality control measures, including regular testing. Proper QC measures also ensure monitoring and control of relevant process parameters, regular inspections of equipment and materials, and performance of thorough QC and release testing at various stages of the cell expansion process.

Manufacturing technology advanced, but improvements needed

The current state of manufacturing technology for cell therapy expansion is generally well-advanced, but according to Tyagi there are many areas where improvements would be beneficial. Many different bioreactors are available, including stirred-tank, rocking-motion, hollow-fiber, and fixed-bed systems, with each having its own set of advantages and disadvantages. As a result, Tyagi notes the choice of bioreactor can significantly impact the quality and quantity of cells produced. He adds that suppliers are striving to improve bioreactor designs to unlock the full potential of cell therapy applications. Tyagi points to fixed-bed bioreactors as a promising technology for scaling adherent cell therapies with higher yields, but because they are completely closed, direct observation and monitoring are challenges.

Cell harvesting and separation from the growth medium after expansion is typically achieved using centrifugation or filtration methods. The current state of cell harvesting and separation technology is fairly advanced, but Tyagi underscores the existence of challenges related to maintaining cell viability and yield during this process. For instance, while gentle and optimized separation techniques such as density-gradient centrifugation are often used to separate cells from growth medium based on their size and density, process parameters such as rotor speed, separation media, and flow rates typically must be optimized for specific cell types. In addition, while the process is suitable for large cell-culture volumes, high shear forces can damage cells, and the process can be lengthy and require significant energy input, according to Tyagi.

Equipment for cell processing (washing, concentrating, and resuspending) and formulation (addition of additives or excipients) is reasonably advanced, according to Tyagi, but these processes can also create challenges with respect to maintaining cell viability and functionality. For instance, closed and automated cell washing and centrifugation-based technologies can efficiently handle large volumes of cells while maintaining their integrity but can be expensive, require skilled operators, are not suitable for all cell types or very large-scale operations, and there are only a limited number of suppliers offering this type of specialized equipment.

With respect to quality control for allogeneic cell therapy expansion processes, there is a need for more rapid and comprehensive testing methods, according to Nanda. She notes that traditional microscopy and cell counting can be time-consuming and may not capture all relevant information about the cells, while real-time monitoring systems, such as in-line sensors or imaging technologies, can provide valuable data for process optimization and product characterization come with considerations like high cost and technical complexity.

Finally, integration and validation of automated and closed systems for cell-therapy manufacturing can be challenging. In addition, while automated cell culture systems can control all aspects of the cell culture process, from seeding cells to harvesting and processing them, Nanda comments that they can be expensive to implement and maintain, and may require significant customization depending on the specific cell-therapy product being manufactured.

Cynthia Challener, PhD is a contributing editor for Pharmaceutical Technology Group.

When referring to this article, please cite it as Challener, C. Challenges to Cell Expansion for Allogenic Cell Therapies. 

Manufacturing

Sourcing of donor cells, access to fit-for-purpose reagents and manufacturing systems, and scalability are top issues.

Challenges to Cell Expansion for Allogenic Cell Therapies 

In today’s biologics/contract research organization (CRO) labs, more data than ever are being generated on protein characterization, thanks to advanced and more thorough analytical technologies. Managing these volumes of data, especially in the clinical phases of drug development, has become a real challenge for researchers.

Clinical trial data are at the heart of how biopharma companies develop new therapeutics for commercial launch, but developing a new drug and bringing it to market can take a decade or more and cost upwards of a billion dollars (1). Outsourcing clinical trial services has become a common practice to save a biopharma company on cost and time, but the practice brings up risks in data transfer as well as data management.

Laboratory issues can result in a delay of data receipt as well as poor quality of data, which can lead to rejection or rework. Data entry issues, delays in responses to data queries, errors during data transfer and merger, and discrepancies due to poor database setup can all also be the cause of delays (1).

A discussion with David Hardy, PhD, senior manager, Data Analytics and AI Enablement, Digital Science Solutions, Thermo Fisher Scientific, explores best practices for how to approach data management in today’s biopharmaceutical lab and highlights some effective strategies for distilling meaningful interpretation from these data.

 Although solutions such as laboratory information management systems (LIMS) and the electronic lab notebook (ELN) have helped to address the vast amounts of data being generated in biologics/CRO labs, what are the biggest challenges facing LIMS and ELN?

 The biggest challenge when it comes to LIMS and ELNs is ensuring that these systems fit seamlessly into laboratory processes to make sure they are used throughout an organization, at which point they can be better used to unlock deeper data-driven insights. Usability can also be a challenge of many LIMS and ELN solutions, but it is critical for company-wide, successful adoption.

Another important challenge is that, while LIMS and ELN can help ensure data quality after data capture, they can do little to rectify errors made through poor quality input data. To help improve data quality at the point of data generation, labs should seek to use automation wherever possible.

 Do automation and digitalization truly offer solutions that allow scientists to analyze these data in a meaningful way, or are there as much cons as there are pros to using such advanced technologies for the data scientist?

 The pros and cons of using automation and digitalization tools to analyze large amounts of data are the same across all applications, including biologics. For example, biologics labs still need to conduct initial validation of their tools and constantly check them, requiring close collaboration between the laboratory and data scientists. However, over time, the pros of advanced digitalization and automation tools—greater accuracy, reproducibility, efficiency, and deeper insight—vastly outweigh any initial cons.

Importantly, even with advanced technologies, scientists face the hurdle of understanding their data and which data fields to use. Then, they need to be clear on which model best suits the problem they’re trying to solve. This isn’t always easy but analytics solutions are striving to make this simpler.

 How has the user (i.e., the scientists) experience been affected or changed by the amount and quality of data being generated and the ability to manage that data?

 By their nature, scientists are data savvy. However, with the sheer amount of data that labs generate, as well as the need to manage these data with advanced technologies, scientists have had to become more tech savvy. For example, most scientists have the knowledge and skills to check and analyze their data using languages such as R, Python, and tools such as KNIME [Konstanz Information Miner]. Given the growing importance of these skills, they are now widely taught to undergraduate students.

Scientists have also embraced ‘big data,’ adopting new technologies to such an extent that big data isn’t the obstacle it once was. Given the increasing importance of technology in handling and processing such data, scientists now commonly work alongside their data science, IT [information technology], and TechOps [technical operations] colleagues to find the optimum and most cost-effective solution, which was less common in the past.

Beyond data capture, storage, and analysis, labs increasingly need to be able to use their data to their full potential. Accordingly, we’ve seen scientists increasingly focus on the FAIR (findable, accessible, interoperable, and reusable) data principles. Data [are] only truly useful if [they meet] these requirements.

 What would you say are some key best practice habits or techniques that scientists can employ to managing these data?

 Scientists can—and should—do several things to maximize their ability to manage these data. First, they should embrace automation, as it’s a scientist’s best friend. With more automation, scientists get free time to focus on more important, value-adding activities.

Scientists should also keep in mind that managing all of these data is a team sport. Data Science, IT, and TechOps teams are there to help, so scientists should leverage their expertise and support to maximize their chances of overcoming data challenges.

Finally, scientists should keep up to date on the latest technologies for managing and analyzing data—the field is advancing very rapidly, and taking your eye off the ball, even briefly, can lead to missed opportunities.

 What are some effective strategies that scientists can use to ensure that they are processing quality lab data and not just noise (are there such strategies)?

 As the saying goes, ‘garbage in, garbage out.’ That is to say that the root of good quality data is good input—good experiment design followed by good execution. Scientists can achieve these in several ways.

Advanced analytical tools generate more data in today’s labs than ever before.

BioPharm International, August 2023

Investigating Outsourcing

Balancing AAVs Versus LLVs

A Look into Biologic Scale-Up Strategies

Streamlining Downstream Processes

Challenges to Cell Expansion for Allogenic Cell Therapies

Dealing with Data: How to Keep It Manageable

The United States and the Global BioPharma Market

Staffing Shortages: Major Hurdle for Bioprocess Contract Manufacturing Services in 2023

Industry Challenges Medicare Price Negotiation Scheme

Supply Chain Securities

New Magic from mRNA

Sharing Critical Information for the Best Outcome