Significant growth in the development and manufacturing of biologics—from traditional recombinant proteins and monoclonal antibodies (mAbs) to advanced multispecifics, antibody-drug conjugates, viral oncolytics, and cell, gene, and gene-editing therapies—was occurring worldwide before the emergence of the COVID-19 pandemic. Demand for biosimilars is also increasing. The requirements associated with COVID-19 vaccine manufacturing are placing further pressure on upstream manufacturing capacities.

“The situation is evolving as the industry continues to explore the right balance between internal capacity/capability and external capacity/capability and key strategic partnerships,” says Lynn Bottone, vice-president of Pfizer Biotech Operations. “Generally, the industry doesn’t have idle capacity across key technologies to bring on line with immediate effect. The challenges of the last year have made the industry revisit and reconsider these options with a rethinking of certain approaches,” she adds. That includes the supply of key inputs to upstream processes as well.

As a result, a strong and sustainable supply network is more vital to ensure supply chain stability, according to Youngsun Kim, director of the manufacturing science and technology team at Samsung Biologics. “Global sourcing of raw materials is an integral part of the supply chain, but recent events have highlighted that relying only on a single source can increase risk during a crisis. Many companies are looking to diversify their supply chains to reduce the risk of disruption in the future,” he says.

Equipment suppliers, contract manufacturers, and biopharmaceutical companies are implementing new bioprocessing solutions and manufacturing strategies to increase efficiency and productivity, forging innovative partnerships, and investing in additional physical capacity on a global basis.

The increasing diversity of advanced therapies has driven upstream manufacturing processes to evolve, according to Philip Vanek, CTO at Gamma Biosciences. “The latest iteration of technologies is being developed with scalability, process intensification, and lean principles in mind to ensure quality and reliability for the life of a process,” he asserts.

Process flexibility and technical innovations are key strategies to accelerate the pace of process development while increasing upstream capacity, adds Dirk Lange, CEO of Selexis and president and CEO of KBI Biopharma. “Accelerating processes means going into manufacturing faster, and with that there can be bottlenecks. CDMOs [contract development and manufacturing organizations] must provide flexibility to address bottlenecks, like taking the most streamlined path possible, decoupling upstream and downstream processes, and managing technical risks more effectively (i.e., using new technologies that help create flexibility),” he explains.

Developing and scaling up manufacturing operations and analytical testing capabilities are, however, requiring major innovation and the courage to move away from conventional constructs, asserts Bottone. “The paradigm shift only occurs with reimagining the way we’ve worked previously, and this reimagining is required at all functional nodes of the product lifecycle and through the end-to-end supply chain. A focus on our horizontal integration internally and externally has been a key enabler for the COVID-19 vaccine and will be key to future new product introductions,” she notes.

The upstream capacity landscape does, however, vary largely with the product class, process design, and technology selection, notes Tania Pereira Chilima, deputy CTO of Univercells Technologies. She points to the mAb sector where stainless-steel bioreactors with tens of thousands of liters of capacity were commonly used for cell culture, but today have been replaced with single-use (SU) bioreactors with thousands of liters of capacity due to significant increases in upstream productivity. The use of perfusion processes has enabled even larger throughputs within the same bioreactor volume.

Furthermore, Chilima observes that by using scalable technologies that have representative scale-down models early in the process development, companies are able to evaluate the capacity needed early on and plan accordingly. Innovation in process analytical technologies (PAT) is similarly driven by the need to boost both the productivity and therapeutic value of each bioprocess, according to Vanek.

Shortening development timelines has been another focus of the industry. “Efforts have targeted the development of new expression technologies, parallel tracking of activities that are not on the critical path, and implementing high-throughput, small-scale technologies,” comments Atul Mohindra, senior director for research and development with Lonza. This work, he adds, has resulted in a significant reduction in process development timelines and is therefore having a positive impact on upstream capacity.

Digital transformation also affords good opportunities to improve lead times for rapid cycle process development, upstream capacity availability, and quality, according to Kim. Samsung, for example, is leveraging automation, ultra scale-down modeling in microreactors, 

 and predictive modeling including metabolomics, and mobile apps for company-wide data visibility to reduce the client tech-transfer process to as little as three and one-half months while maintaining high levels of quality.

Viral vectors, which are used directly in therapeutics and vaccines and also in the production of gene and gene-modified cell therapies, represent a special case with respect to manufacturing capacity shortages.

Traditionally, viral vectors were manufacturing using plasticware such as roller bottles and T-flasks and manually-intensive processes that required scale out rather than scale up to increase capacity, leading to severe constraints, according to Chilima. “These challenges can be overcome through the use of scalable technologies such as stirred-tank bioreactors or even intensified, scalable technologies such as fixed-bed bioreactors,” she says.

Upstream capacity in viral-vector manufacturing is also impacted by capacity limitations in downstream processing. This issue is currently leading many CDMOs to reject projects or set long wait times for new customers to ensure they are able to fulfill their existing contracts, even though they may have under-utilized upstream capacity, says Kai Lipinski, chief scientific officer at Vibalogics.

On a positive note, parallelization of experiments using automated systems for high-producer clone selection, along with medium optimization and the availability of standard kits, are having an overwhelmingly positive impact on capacity, according to Lipinski. The implementation of platform processes that require no or minimal development (e.g., a single cell line is used for many products along with a platform production medium and optimized vector platform design) is also saving time and cost. Similarly, accurate scale-down models and the use of a quality-by-design approach during development are minimizing risk and providing increased predictability when moving to large-scale production

The one factor currently limiting the pace of change, Lipinski asserts, is talent availability. “More expertise is needed across the industry to help companies identify new ways to further improve process development to make better use of existing upstream capacity,” he states.

The COVID-19 pandemic has created greater interest and awareness in the biotech sector, which seems to be leading to more investment and greater utilization of current biologics manufacturing capacity, says Jeetendra Vaghjiani, director of commercial mammalian development at Lonza. Overall, though, he notes that COVID-19-related projects including vaccines are using up capacity that may have been used for other therapeutic products, which has increased the demand for upstream capacity.

Tony Hitchcock, technical director for Cobra Biologics, disagrees. “The production of COVID-19 vaccines and therapies is largely disconnected from traditional biologics as the first authorized candidates are based on adenoviral technology (AstraZeneca/Oxford and Janssen) or messenger RNA (mRNA) (Pfizer-BioNTech, Moderna), with candidates still in development being DNA-based (Scancell), protein-based (Novavax), and using more classical technology (Valneva). This manufacturing is performed outside of mainstream biopharma, where most capacities and capabilities involve the production of mAbs,” he explains.

While Lange says that COVID-19 has created a temporary shortage of upstream capacity for a portion of the industry, this spike has been counteracted by a stronger focus on life sciences as a whole, with more funds being pushed into biotech for other vaccines and therapeutics. “Large capital investments will provide relief and support growth of the industry in the mid-term,” he remarks.

Biopharmaceutical companies of all sizes have sought additional contract manufacturing capacity to free internal capacity, expedite process development and large-scale manufacture, and forge partnerships to secure secondary sources in their drug supply chains, according to Kim. “Having a stable secondary source plays an essential role when it comes to supply continuity, especially during times of uncertainty like COVID-19,” he states.

For instance, Regeneron has been working with FDA and the US government to rapidly scale up production of its REGEN-COV antibody cocktail. The company is leveraging production and manufacturing platforms developed over decades, financial support from BARDA, and accelerated licensing of its Irish facility.

“This approach has enabled us to adjust much of our internal manufacturing activities to maximize our ability to produce REGEN-COV while still ensuring patients get the other FDA-approved Regeneron medicines that they need,” says Hannah Kwagh, senior manager of corporate communications at Regeneron. The full portfolio of commercial products can be manufactured in Ireland, enabling the company’s US site to maximize production of REGEN-COV.

“Right now, under our agreement with the US government, we expect to deliver approximately 750,000 doses by June 30, 2021, with the vast majority of these deliveries occurring in the second quarter. We are evaluating a lower dose (1.2 gram) for REGEN-COV and should this lower treatment dose receive Emergency Use Authorization or approval from the FDA, we aim to deliver up to 1.25 million doses by June 30, which is the maximum quantity that is authorized for purchase by the US government under this supply agreement,” Kwagh says.

To support its COVID-19 vaccine (BNT162b2), Pfizer had to both expand its capability and build new capacity for the upstream process, which it did in part through various partnerships. “Industry collaboration is evident in numerous examples where companies have supported one another to get a vaccine or a therapeutic to patients faster. The industry has been pretty courageous not only in expanding or building capacity without knowing whether the platform/product would be successful, but also in extending unutilized or underutilized capacity to competitors for the common good,” states Bottone.

There have, however, been supply chain challenges attributed to the COVID-19 pandemic. “Vendors have rapidly shifted focus to produce materials to support COVID-19 vaccines and therapies, and as a result some therapeutic developers have had to slow down or redirect demand for equipment, reagents, and consumables,” says Vanek.

In general, adds Christian Morello, head of global supply-chain planning and programs management at Lonza, the demand for materials used for the manufacture of biological products has increased over the past five years, and mainly for SU equipment, and the accelerated development and production of COVID-19 vaccines has increased the demand much faster than suppliers have been able to increase capacity.

“The upstream processing industry, especially in the SU space, was already experiencing significant demand and growth pre-COVID-19. With the rapid development of new COVID-19 vaccines and therapeutics, incremental capacity for both hardware and consumables is coming on line at a record pace. That said, there are challenges within the supply chain as the industry addresses shortages in raw material and component availability, affecting overall lead times in many areas,” agrees Jon Van Pelt, general manager of Cytiva’s bioprocess, single-use, and enterprise solutions business. He anticipates continued supply constraints in certain areas for the next several quarters as new capacity comes on line.

“The constrained supply chain for such materials is creating the need for daily management and frequent reshaping of production plans, while also accelerating the validation of dual sourcing to decrease exposure to such risk,” Morello observes.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak has highlighted the need for high-capacity, scalable technologies for viral vaccine production, adds Chilima. “This outbreak has shown that using manual technologies is not a suitable strategy when large quantities of doses are required during a pandemic. Many companies using adherent cells have been driven to consider more scalable technologies that maintain a similar microenvironment for the cells,” she notes.

On the positive side, Hitchcock notes that the COVID-19 pandemic is not only stimulating new investments from the private sector and strategic investments from governments to establish local vaccine production capabilities, but also accelerating the development of these technology platforms and their broader application beyond cancer indications.

Contract manufacturers, drug developers both affected

Both biopharmaceutical companies and their outsourcing partners have both been impacted by the increased demand for upstream bioprocessing capacity. “This increased demand impacts equally classical pharmaceutical capacity and CDMO capacity as it is the product that is important,” Morello says. “In that perspective,” he adds, “it was interesting to see the collaborative approach on the supply chain between sponsors and CDMOs.”

KBI Biopharma has observed that clients focused on COVID-19 vaccines have limited in-house capacity. As a result, many are moving other vaccine products with shorter timelines to CDMOs, leading to the increased demand across the industry as all manufacturers adapt to the manufacturing challenges created by the pandemic while still maintaining production of other vaccines and therapeutics.

Vaccine manufacturing in terms of capacity, according to Hitchcock, has largely been provided by CDMOs operating in the cell and gene therapy space for viral vector and plasmid DNA production. “Prior to the pandemic, large investments in capacity were being put in place with the likes of Cobra Biologics in the UK completing its viral vector production expansion in 2020 and with other significant expansions being carried out globally to keep up with the growth of the advance therapies sector. This activity was certainly serendipitous,” he comments.

Lipinski believes, however, that CDMOs are likely to be more impacted than biopharma companies by upstream capacity issues because they cannot easily divert capacity committed to existing customer projects to COVID-related projects without authorization from those customers. “Drug developers, on the other hand, have greater freedom to define and change their development priorities, allowing them to refocus capacity on the fight against the pandemic,” he remarks.

That is at least true for large biopharmaceutical companies, but small and emerging biotech companies typically rely on the manufacturing capacity, expertise, and flexibility of CDMOs to bring their candidates to market. “With most of the drug development pipeline being held by these emerging and small biotechs, CDMO bioprocessing capacity is gaining importance,” Vaghjiani states.

Despite the challenges, Lipinski does stress that CDMOs have quickly responded to support customers seeking to deliver new COVID vaccines and therapies.

Noticeable shifts in manufacturing strategies

While the addition of more upstream capacity has been occurring and continues to take place, CDMOs and biopharmaceutical companies are also investing in new technologies and manufacturing strategies to achieve capacity increases.

“There is drastic disruption needed to support the commercialization of advanced therapies,” Vanek asserts. He points to three emerging themes that are dramatically improving manufacturing efficiency across the biologics spectrum and thus ultimately impacting upstream bioprocessing capacity: continuous manufacturing, process analytics, and radically improved downstream purification. “There is a real shift,” agrees Chilima, “towards intensified technologies that enable dramatic reductions in operational footprints and increased overall productivity.”

There is also a lot of focus on compressing the biologic drug development timeline within the industry, especially in the process-development space, according to Van Pelt. “If you think broadly, from process development to clinical to CGMP [current good manufacturing practice] production, advances in digital solutions are accelerating the pace of drug development. Validated platform processes, modeling, connectivity, PAT, and other digital tools will allow for increased speed and process understanding while reducing the risk of failure as the process advances through the various development stages,” he observes.

Selexis and KBI Biopharma, for instance, have invested in technology and innovation in cell-line development and process development to reduce cost of goods while increasing output at a given scale. High-yielding cell lines and process intensification are powerful strategies to address growing demand, according to Lange. The company has also invested in digitalization and manufacturing 4.0 technologies to maximize the effectiveness of its operations.

Lonza, too, has invested in technologies and processes to reduce development, tech-transfer, and batch-release timelines for increased throughput and therefore additional capacity, according to Mohindra. “In addition, we are further improving right-first-time success rates and our lean management systems and have implemented in-line testing technologies that leverage digitalization to increase productivity,” he says.

Pfizer has leveraged production systems and its management infrastructure to free up time for continuous improvement of operations and innovation, according to Bottone. “Expanding our digital capabilities has been important in the pursuit of big data analytics that has enabled efficient and effective development, analysis, and review of significant quantities of data. Automation of historically manual processes has been equally impactful,” she notes.

Development of rapid analytical methods; the use of robust inline, online, and at-line monitoring and testing capabilities to reduce the reliance on end-product testing, exploration of accelerated regulatory pathways to approval for emerging technologies, application of the use of digital twins to increase operational efficiencies, and the application of artificial intelligence to historically manual processes like automated visual inspection are all taking place at Pfizer.

Innovation in storage and distribution was evident with the mRNA vaccine roll-out, Bottone adds. “Modular and mobile solutions will be important for scale-up and capacity-expansion opportunities with much shorter lead times,” she observes.

SU technologies are also being widely implemented for commercial as well as clinical production to minimize the time and costs associated with maintenance and cleaning of standard reusable equipment, according to Lipinski. SU bioreactors are now available up to 5000 L, enabling larger batch sizes. Companies are also leveraging high-cell-density perfusion processes to boost efficiency, according to Lipinski.

Increased focus on alternate sourcing for both materials and intermediates and use of automation, production scheduling, and batch release are becoming more prominent, comments Kim. “The supply of raw materials and consumables is critical to ensuring the timeline for large-scale production, especially at the moment. What we’re seeing across the board is sponsors providing alternative (secondary and tertiary) vendors for key raw materials and CDMOs taking responsibility for establishing an alternative consumable inventory list for faster procurement, especially for the commonly used consumables in the biopharma industry,” he observes.

Means for enhancing viral-vector productivities are also being explored, such as screening for high-producer cell clones, using technologies to achieve high cell densities and perfusion cultures, and optimizing virus recovery. In addition, Lipinski notes that the use of high-efficacy viral vector platforms to minimize the needed vaccination dose can play an important role in helping to make better use of existing capacity.

On the front end, the use of prefabricated container-like clean rooms in new facilities is accelerating timelines for the completion of capacity expansions, according to Lipinski.

Both CDMOs and biopharmaceutical companies are investing heavily in long-term capacity, with individual investments too numerous to list. KBI Biopharma, for instance, is implementing a global expansion strategy that includes bringing online new clinical and commercial manufacturing facilities in the United States and Europe.Vibalogics is in the middle of a $150-million investment at its site in Boxborough, MA, and is currently also investing extensively in its early-phase facility in Cuxhaven, Germany.

Samsung implemented large-scale N-1 perfusion to enable inoculation of production bioreactors (15,000 L) at higher cell densities and achieve peak cell densities within shorter culture durations. The company has also invested in process automation, including a manufacturing execution system, and online monitoring capability and is looking to fully automate its activities. A new manufacturing facility is also under construction and expected to be operational by the end of 2023. “The newly built plant, which will be the world’s biopharmaceutical manufacturing facility, will be fully equipped with multi-scale bioreactors and advanced technology to meet the trending needs of the market and clients,” Kim says.

Significant activity has occurred in the plasmid DNA and viral vector space. Cobra Biologics, which was acquired by Cognate Bioservices in 2020 and is soon to be acquired by Charles River Laboratories for $875 million (1), is quadrupling its clinical and commercial capacity for high-quality DNA manufacturing in Europe and doubling its capacity in the US. Some other companies that have recently completed or announced new capacity include Matica Biotechnology, Porton Biologics, Fujifilm Diosynth Biotechnologies, Oxford Biomedica, Emergent BioSolutions, BioReliance (Merck KgaA), WuXI Advanced Therapies, Delphi Genetics, Biomay, VGXI, and Thermo Fisher Scientific.

SU equipment suppliers have also responded to the need to increase output and reduce lead times for disposable biopharmaceutical manufacturing solutions. Pall Corporation, for instance, is expanding capacity at six existing manufacturing sites in Europe and the US and constructing a new manufacturing facility in the US (2).

The need for access to greater upstream (and downstream) bioprocessing capacity has led to the formation of interesting partnerships across the industry. “The current crisis has triggered a need for companies to collaborate even more closely and think out of the box to address issues beyond company boundaries or legal entities. This virus doesn’t respect corporate or geopolitical boundaries, so the industry has had to respond accordingly,” Lange says.

Because adding new capacity can take time, the current need for immediate capacity is leading companies to find innovative ways to meet capacity demands, adds Vaghjiani. Partnerships and collaborations are an important strategy for expediting capacity expansion plans to ensure there is enough upstream bioprocess capacity to manufacture the vaccines and therapeutics needed to address the COVID-19 pandemic, agrees Oliver Loeillot, vice-president of Cytiva’s bioprocess business. “Depending on the needs of each organization, partnerships and collaborations will remain a strategy for capacity expansion for the foreseeable future,” he says.

“Partnerships between biopharma companies will provide flexible and agile supply options driven by demand and capacity,” asserts Kim. “Increased partnerships across all supply chain segments will de-risk supply and the overall required investment and give access to flexible manufacturing capacity supported by innovative technologies and facilities, extremely well-trained staff, and competitive pricing,” he continues.

Numerous industry collaborations, including strategic partnerships between companies and contract service providers exploring ideas around flexible capacity and partnerships with vendors and suppliers to better understand and solve their capacity challenges, are playing a role alongside significant capital investments in capacity and capability and a paradigm shift in reducing capital project timelines to support capacity expansion, Bottone adds.

For Lipinski, such partnerships are crucial to increasing production scales within a reasonable time frame and to addressing upstream biopharma capacity needs in the future. “Small and newly-established biotech companies often have little to no drug development or regulatory experience, meaning they need supportive partners with the knowledge and capability to bring their innovations to market. The same is true for supply-chain management of drugs,” he says.

Large pharmaceutical companies, with their well-established experience in biopharma development and regulatory insight, can help these smaller partners, Lipinski observes. “They can play a key role in supporting and minimizing development timelines for Phase I/II/III all the way through to commercial.” He also notes that CDMOs are also well placed to provide this support, delivering infrastructure and regulatory expertise to address gaps in small biotech companies’ knowledge.

Manufacturers will also need to work very closely with vendor partners to project longer-term supply needs and evaluate new alternatives for materials that are most impacted by the vaccine manufacturing demand, says Lange.

For Regeneron, innovative partnerships have been key to ensuring that the company is getting medicines and vaccines to patients as efficiently as possible, according to Kwagh. For example, she notes that while Regeneron currently anticipates that the full supply of REGEN-COV for its agreement with the US government will be produced by Regeneron, the company has also significantly increased global capacity through a collaboration with Roche. “Collectively, we now expect to be able to produce more than 2 million treatment doses per year, and if data supports use of a lower dose of the cocktail this figure would increase accordingly,” she says.

“The whole life sciences industry is coming together to address this global crisis and moving forward, we hope this will inspire collaboration to achieve the common goal we share—delivering life-saving treatments faster than ever to the patients who need them most—every day, not just during a crisis,” concludes Lange. 

Addressing bioprocess capacity issues is essential to the future success of many biopharmaceutical products. Regeneron’s ability to provide high numbers of finished doses of REGEN-COV is predicated on continued success in the manufacture of bulk product and access to third-party fill/finish capacity, which is under heavy demand from COVID-19 vaccine manufacturers, is just one example. “Looking ahead, as an industry we need investment in both R&D technology and manufacturing infrastructure in order to better prepare for future pandemics,” Kwagh says.

The innovative collaborations/relationships that have been established during the pandemic should continue to be explored as key strategic levers, notes Bottone. “The idea of flexible capacity that can be brought on line with immediate effect is an interesting and appealing proposition when you consider our collective experiences of the last year with the pandemic,” she says. In addition, Bottone suggests that collaboration between clinicians, regulators, and industry will determine how much acceleration of drug development and commercialization is possible, and the signs are encouraging.

The COVID-19 pandemic, according to Lange, is a case study in what happens if global capacity models are unexpectedly disrupted. Fortunately, many innovative technologies are being developed with the potential to enhance and shape the future of biomanufacturing, Vanek observes. “The challenge lies in scaling these technologies in ways that will unleash productivity and spur further innovation,” he states.

Across the industry, though, Vanek believes there has been a gap in investment capital for companies that have launched these innovative products but still require guidance to fully scale and industrialize their solutions. “Without expert guidance and proper representation. these innovations could be lost or stifled if they are acquired or merged into large life sciences companies with many competing priorities,” he says.

The supply of crucial SU upstream process equipment and consumables is a key concern for Lipinski, with delivery times of up to 52 weeks currently expected across the industry. He also notes that aseptic filling of biologics, in particular live virus filling, is one of the most critical bottlenecks in the biopharma development and manufacturing process, including availability of the required primary packaging material. This bottleneck ultimately impacts upstream capacity too.

“Steps need to be taken to address these issues as quickly as possible in order to optimize biologics manufacturing capacity,” Lipinski says. He adds that the development of a form of generic consumable equipment and packaging in the near future may help to overcome the overall shortages of key materials, allowing the increased production of viral vector and virotherapy products using SU production systems.

In the meantime, companies developing new biologics, and in particular vaccines, oncolytic viruses, and gene therapies based on viral vectors, need to identify and access manufacturing capacity as early as possible to ensure they can bring their innovations to market as soon as they receive regulatory approval, according to Lipinski. “Waiting for limited capacity to become available is not practical, and the earlier they engage with a potential manufacturing partner, the more control they will have over their timelines and speed to trial,” he asserts.

Access to talent is another issue that must be addressed to ensure future success in biomanufacturing. “Every fast-growing company has experienced the limiting effects of a defined talent pool and an overall shortage of qualified personnel. Capital investments, engineering projects and process improvement are fundamental to enable the projected growth of our industry, but we also need to work on a sustainable pipeline of talent at all levels in our organizations,” Lange stresses.

KBI Biopharma is tackling this problem by working closely with educational institutions and large industry networks to create a sustainable workforce to support its growth. Lipinski observes that the need to ensure a workforce with the necessary depth of expertise is in part also driving larger biopharmaceutical companies to bring technology, capacity, and talent inhouse via acquisitions.

In general, summarized Bottone, access to key data in terms of capacity and capability is important for the biopharmaceutical industry as a whole, and leveraging these data to understand the current and potential challenges for all nodes of the supply chain, upstream and downstream, will facilitate the prioritization of near-term and long-term action. “The COVID-19 vaccine has paved the way for paradigm shifting innovation in accelerating new product introductions, enabling the industry to deliver breakthroughs that change patients’ lives,” she concludes.

1. Cobra Biologics, “Charles River to Acquire Cognate BioServices to Create a Premier Scientific Partner for Cell and Gene Therapy Development,” Press Release, Feb. 17, 2021.
2. Pall Corporation, “Pall Corporation Expands Production Capacity in the United States and Europe to Support Industry Demand,” Press Release, Jan. 7, 2021.

Cynthia A. Challener, PhD, is a contributing editor to 


Vol. 34, No. 4
April 2021
Pages: 10–17

When citing this article, please refer to it as C. Challener, "Maximum Output Starts with Optimized Upstream Processing," 

Articles

Cover Story

Biopharmaceutical companies and contract manufacturers respond to changing demand dynamics for upstream bioprocessing capacity.

Maximum Output Starts with Optimized Upstream Processing

In this article, the authors have detailed the best practices when optimizing and sizing anion exchange (AEX) membrane adsorbers for virus clearance. AEX chromatography is a powerful method to clear all types of virus by separating viruses from the product of interest based on ionic charge. Membrane adsorbers (macroporous membranes with chromatographic ligands attached that function similarly to resins containing identical ligands) offer several advantages over conventional resin chromatography, such as higher binding capacity for large molecules (i.e., viruses), >10-fold faster flow rates, use of up to 75% less buffer, and present a flexible single-use solution that does not require cleaning validation. These qualities make membrane adsorbers a perfect fit for flow through chromatography operations where virus clearance is expected. The authors therefore performed minute virus of mice (MVM) spiking studies with a monoclonal antibody product and small-scale AEX membrane adsorbers to evaluate a range of critical processing conditions, such as conductivity, process pause, product loading, and virus spike purity. The results provide an understanding of the impact of the evaluated process parameters on the clearance of parvovirus.

Submitted: Nov. 5, 2020.
Accepted: Jan. 20, 2021.

Sherri Dolan is a global technology consultant for Virus Clearance at Sartorius North America; Paul Nolan and Craig Quinney are process scientists; Stephen Carey is an analyst; and Lorna Littlejohn*, lorna.littlejohn@sartorius.com, is the manager of Confidence Virus Clearance Services; all are at Sartorius in Glasgow, Scotland.

*To whom all correspondence should be addressed.


Vol. 34, No. 4
April 2021
Pages: 32–37

When referring to this article, please cite it as S. Dolan, et al., “Using Anion Exchange Membrane Adsorbers to Ensure Effective Virus Clearance of Challenging Parvoviruses,” 

Peer-Reviewed Research

A series of minute virus of mice spiking studies were performed with a monoclonal antibody product and small-scale AEX membrane adsorbers to evaluate a range of critical processing conditions.

Using Anion Exchange Membrane Adsorbers to Ensure Effective Virus Clearance of Challenging Parvoviruses

The global appetite for biological therapeutics is growing, with estimates for the market to surpass US $4 billion by 2025 (1). For the most part, biologics are delivered intravenously as a result of the complex and large nature of biomolecules, poor permeability through the intestinal epithelium, and susceptibility to enzymatic degradation in the gastrointestinal tract (2).

However, injections are considered to be painful and inconvenient by many patients, particularly in cases where chronic conditions are being treated. Therefore, alternative routes of delivery, such as inhalation, for biologics are being investigated to improve systemic drug absorption and overcome the treatment burden associated with intravenous administration (3).

To learn more about inhaled administration of biologics—the advantages of this delivery route as well as its limitations and challenges—

 spoke with Catherine Oliver, specialist—Pharmaceutical Development, Vectura.

 Why is inhaled administration of biologics attractive when compared with other delivery routes?

 Traditionally, biologics are given through injections or infusions due to poor oral bioavailability. However, up to one in 10 of the population are anxious about injections. Administration of infusions requires skilled personnel to prepare and administer.

Drug delivery via inhalation offers obvious advantages for treating diseases of the lung. For respiratory therapies, it allows the drug to reach the site of action directly. Inhalation can also be a viable route for the delivery of systemically acting drugs, not least because it gives a fast onset of action through high systemic bioavailability.

There is renewed interest in inhalation as a route for administration of biologics, both systemic and locally acting. The COVID-19 pandemic has highlighted the difficulties associated with lung damage, and may well prove to be a catalyst for greater efforts for innovation in this field.

 What are the main obstacles to a successful development program of an inhaled biologic?

 There are two important aspects to consider when developing an inhaled biologic: making particles/droplets that are sufficiently small to be inhaled; and the aerosolization of dry powders produced by spray-drying, lyophilization, or nebulization of solutions for inhalation. Both have the potential to affect product quality through decreased activity or protein aggregation. Biologics tend to be complex large molecules and, as such, have more potential sites for degradation. It is essential that the formulation and packaging system are optimized with the intended delivery system in mind. Therefore, critical quality attributes (CQAs) for the drug product are carefully defined and assessed.

Lastly, patient factors must be considered. Any novel inhaler must have noticeable beneficial impact on therapy effectiveness, compliance and patient lifestyle and be able to demonstrate that it is just as effective at delivering the required dose as other forms of administration.

 Have there been any technological advances that have helped developers overcome the hurdles associated with inhaled biologic development?

 However carefully a biologic has been designed and developed, there is still no guarantee that a successful inhaled formulation will be possible.

A greater understanding of factors affecting biologic stability has resulted in improved drying techniques to produce inhalable dry powders. For example, for biologics that are unstable as aqueous solutions, understanding both the optimal residual moisture content and excipient characteristics can inform formulation for drying processes to ensure a stable dry powder. This can enable room temperature storage and negate the need for cold chain storage.

It has been known for some time that nanoparticles/nanobodies are effective at overcoming pulmonary clearance by mucocilary escalator or macrophage capture. This technology has never been more topical due to clinical trials of COVID vaccines, containing mRNA [messenger RNA] encapsulated in lipid nanoparticles, being administered by nebulization.

Whilst there have been successful biologic dry powder inhalers (DPIs) delivery of a liquid formulation using some form of nebulizer is more likely to be considered. The innovator now has a choice of nebulizer types for precise delivery to specific areas

of the lung, such as the use of smart nebulizers to deliver anti-infective treatments deep into the lungs. New DPI and nebulizer inhaler technologies are being developed to facilitate proper inhalation technique by providing feedback during drug administration to the patient or the health-care professional during inhalation.

 Do you believe that the limited number of ‘approved’ excipients for use in inhaled biologics poses a significant challenge to development?

 This is certainly a challenge. The number of excipients for parenteral biologics is about 300; however, less than 20% of these are permitted for inhalation. Historically, the majority of excipients have been approved to support formulation for oral drug delivery. Increased scientific understanding is driving targeting of drugs either to specific parts of the lungs or sites within the body. Both biologics and other new inhaled medicines may need new or innovative technologies to succeed.

There may be times when gaining approval for a novel excipient is necessary because a novel-product-enabling approach is being taken. The need for new excipients will have to be assessed on a case-by-case basis, such as where the stability of the formulation is sub-optimal, or its flow properties cannot be made acceptable with existing substances.

One area where an excipient could be advantageous is in increasing the residual time of the drug in the lungs, rather than it being absorbed and transported. This is something of a holy grail for developers, but if the aim of a novel inhaled formulation is systemic delivery, the opposite is required: very rapid uptake. The potential for a novel excipient in this area here, too, is large.

 Can reformulation provide a convenient route for developers to achieve an inhaled formulation of a biologic?

 Inhalation may be a potential route for a successful reformulation of a marketed biologic, particularly if faster onset of action might be beneficial. However, it does not always follow that an active ingredient that is stable in a liquid formulation for intravenous delivery can demonstrate stability when delivered by inhalation.

It is possible that an existing formulation can be nebulized; however, it is more likely that it will require reformulation. The indication will impact the choice of the delivery method/device to be used. This will then dictate the presentation and formulation choices. Each presents a novel challenge: stability as a dry powder formulation in a moisture-free environment versus sensitivity of liquid formulations, to shear forces during nebulization. This presents an opportunity to simplify the existing formulation, for example, to remove excipients, such as buffer salts or non-reducing sugars used to stabilize spray-dried/lyophilized proteins.

Creating a suitable formulation for the chosen delivery system will require an integrated approach between analytical scientists, process experts, and device engineers to be successful.

 Are there any regulatory challenges that developers should consider when approaching inhaled biologics?

 There are significant regulatory challenges in this area, and with so few approved inhaled biologics published, FDA guidance is limited. The FDA guidance for metered dose inhalers and DPIs used for small-molecule drugs was updated and a new draft issued in April 2018, while the guidance on ‘nasal spray and inhalation solution, suspension, and spray drug products’ was finalized in July 2002 (4,5).

The inhalation field continues to progress significantly. Industry groups, such as The International Pharmaceutical Aerosol Consortium on Regulation & Science and European Pharmaceutical Aerosol Group, which Vectura supports, are working to establish best practice for innovators in the development and registration of inhaled biologics.

1. BCC Research, “Biologic Therapeutic Drugs: Technologies and Global Markets,” Market Research Report (April 2021).
2. W. Liang, et al., 

, 12 (11) 1025 (2020).
3. J.O. Morales, et al., 

, 19 (May) 652–668 (2017) [and references therein].
4. FDA, 

Metered Dose Inhaler (MDI) and Dry Powder Inhaler (DPI) Drug Products—Quality Considerations

, Guidance for Industry, Draft (CBER, April 2018).
5. FDA, 

Nasal Spray and Inhalation Solution, Suspension, and Spray Drug Products—Chemistry, Manufacturing, and Controls Documentation

, Guidance for Industry, (CBER, July 2002).

Felicity Thomas is the European editor for 


Vol. 34, No. 4
April 2021
Pages: 18–20, 46

When referring to this article, please cite it as F. Thomas, “Taking a Breath: Advances in Inhaled Biologics,” 

Development

The inhaled route of administration for biologics is experiencing renewed interest, particularly in light of the COVID-19 pandemic.

Taking a Breath: Advances in Inhaled Biologics

Containment of ingredients and materials during bioprocessing is necessary for the protection of the manufacturing environment, the operators working in that environment, and the drug product. Because containment is an ongoing priority for the industry, innovative technologies are needed to support it. For example, as the adoption of single-use systems in bioprocessing grows, the introduction of newer single-use technologies to compliment those systems are helping to streamline aseptic bioprocessing. For instance, single-use bags for transferring powder ingredients (e.g., cell culture media) to the bioreactor are an increasingly useful tool in keeping the loading of the bioreactor a closed process.

It has been historically accepted that sterility is not a critical factor early on in the biomanufacturing process, such as during the transfer of cell culture ingredients, for instance, powder ingredients for media and buffers. Thus, powder ingredients have traditionally been scooped from stock containers and then weighed and mixed in vessels or bags, a process that is typically done in a separate clean room to control contamination from airborne material (1).

However, as biomanufacturers grow to use single-use technologies, single-use bags can also be applied for other tasks besides the cell culture process itself. Single-use bags offer a simpler, cleaner, and safer way to handle ingredients. Dry ingredients can be transferred to single-use bags with the added convenience of having pre-weighed amounts prepared from storage vessels.

To discuss the challenges and practicalities of using single-use bags for handling powder ingredients in bioprocessing, 

 spoke with Ben Wylie, senior product manager, ChargePoint Technology, a provider of high containment and sterile powder transfer solutions.

 Why is the development of single-use bags for powder handling/powder transfer considered an impactful innovation in upstream bioprocessing? What is their function; how do they work?

 The rise in demand for biopharma treatments in recent years has led to a significant increase in the need for aseptic manufacturing. Biologic treatments are particularly reliant on sterile processing due to the fact that they are most often administered parenterally to patients, so additional steps must be taken to ensure they remain safe.

A major challenge facing pharmaceutical companies looking to meet this new market demand for sterile treatments is how to achieve an aseptic processing environment at every stage in production without compromising on efficiency and productivity. For a growing number of drug developers and their contract development and manufacturing organization (CDMO) partners, the answer to this conundrum is the use of single-use (SU) technologies in their processing, such as single-use bags for handling and transferring powders.

Single-use powder transfer bags are made from a high purity polyethylene (PE) film liner and are specifically designed for use in pharmaceutical and biotech applications. They are utilized in bioprocesses for the safe and efficient transfer of media powders and for buffer powders. Downstream, in fill/finish applications, the bag forms part of a complete closed transfer system for the sterile transfer of API powders to the formulation process.

 What ‘old school’ challenges or issues do these single-use bags address and/or fix for biomanufacturers?

 This new generation of SU equipment is a significant innovation in pharmaceutical development, addressing a key challenge posed by standard multi-use containers and other components—that of production downtime for washdown.

As they are designed to be used once prior to disposal, SU bags and other equipment do not have to be included in cleaning and washdown procedures between batches. All that is required is for them to be replaced with new components. This can significantly streamline the hygiene procedures needed to maintain sterility compared with reusable alternatives, reducing production downtime and enhancing efficiency.

All of this means that SU bags and other equipment can ensure the aseptic integrity of products during transfer within and between manufacturing facilities. They can maintain optimum product quality, while helping to maximize process efficiency and reduce production costs.

 How do these bags play a role in maintaining aseptic conditions for bioprocessing?

 SU bags and similar SU components are manufactured within an ISO6 cleanroom environment and are gamma sterilized prior to use when required. As such, they are suitable for use in the most rigorous aseptic processing environments.

SU bags can be designed to connect to the disposable passive component of a single-use valve, (e.g., Split Butterfly Valve [SBV], ChargePoint Technology) to enable the contained and sterile transfer of pharmaceutical powders between each step in the manufacturing process, as well as between facilities. The passive half doesn’t need to be cleaned after use because it can simply be disposed between fillings, increasing productivity.

Traditional multi-use containers and other components, on the other hand, would have to be washed thoroughly between batches, alongside the washdown of the rest of the line, increasing downtime and the time and headaches often associated with validating cleaning procedures. SU bags help to minimize washdown requirements, reducing production downtime. As a result, they maintain the highest standards of aseptic processing while optimizing manufacturing efficiency.

 Does switching to these single-use powder-handling bags pose their own challenges, and if so, what are they?

 Generally, the time and complexity in adoption of any new single use component lies in the up-front work to qualify the materials and build a robust supply chain. However, this effort pays dividends in the long run with clear benefits in being able to deploy pre-qualified solution off-the-shelf as required. Typically, this solution is easy to retro-fit to an existing process and requires minimal training for line operators to be able to handle and fit.

With all of this in mind, single use components present a highly effective alternative for manufacturers looking to enhance the efficiency of their GMP [good manufacturing practice] and even aseptic powder handling processes.

 Can you explain the cost and overall operating benefits that single-use powder-handling bags offer (if not already addressed)?

 As explained in previous questions, SU bags and other disposable equipment can significantly improve manufacturing efficiency during aseptic processing by reducing the downtime required for washdown and hygiene processes between product batches. By using such components, manufacturers can optimize output on their lines, maximizing manufacturing capacity without compromising on product quality.

1. C. Rombach, “Facing the Challenges: Single-Use Approaches to Powder Transfer,” ILC Dover White Paper, 

Feliza Mirasol is the science editor for 


Vol. 34, No. 4
April 2021
Pages: 22–23

When referring to this article, please cite it as F. Mirasol, “Single-Use Bags Help Maintain Aseptic Ingredient Transfer to Bioreactors,” 

Upstream Processing

The innovation of single-use technologies is applicable early on in the manufacturing process.

Single-Use Bags Help Maintain Aseptic Ingredient Transfer to Bioreactors

The growing use of single-use technologies in downstream bioprocessing allows for opportunities to address problematic areas in unit operations and develop systems to alleviate stresses on the biomanufacturing cycle. One area benefitting from the use of single-use systems is liquid handling and fluid transfer. Single-use components such as tubing, connectors, and valves offer more efficient workflows, for example.

To discuss the challenges and benefits of having a well-designed and properly composed fluid handling system using single-use technology, 

 spoke with Tim Korwan, director, New Product Introduction—Single-Use, at Avantor.

 How have single-use technologies benefitted fluid handling/fluid transfer in downstream bioprocessing? Are there any specific technical challenges that single-use technologies are better able to address and/or resolve?

 In general, single-use equipment gives manufacturers the power to produce more of their product or units/year, and fluid transfer steps in downstream processing can benefit from the implementation of single-use systems. Single-use tubing, connectors, and valve components are among the components that can eliminate cleaning-in-place and sterilizing-in-place (CIP/SIP) procedures, removing these costly and time-consuming steps that contribute to downtime and, thereby, increasing manufacturing throughput and productivity of the facility. Time and personnel once used for CIP/SIP of stainless-steel process lines can now be reallocated to production, increasing the overall efficiency of the manufacturing process.

 What is hose assembly and why is it important to do correctly?

 Building hose assemblies goes beyond measuring from point A to B. Understanding the proper process flow rates to ensure that the fluid pathway meets the specification takes process knowledge and competent knowledge of fluid dynamics. In addition, the single-use system manufacturer must have deep industry knowledge of all the available fittings to be able to ensure that the proper fittings are selected for the particular application and that they integrate well with other components in the system. The selection of the material that will be in contact with the fluid pathways is important from a regulatory and quality standpoint. Finally, the production environment and any pre-use integrity testing will ensure that the hose assembly as designed will deliver the intended result.

 In terms of design aspects, what makes for an efficient hose assembly, and how does this benefit downstream processing steps like chromatography, filtration, etc.?

 Quality-by-design (QbD) in single-use hose assemblies is critical to ensure that the product is designed to meet the specification and application requirements for downstream processing.There has been an increasing demand to use single-use systems for downstream unit operations; two common areas where it is being applied include chromatography and filtration.

Both chromatography and filtration operations are subject to rapid fluctuations in pressure. Rapid changes in pressure can compromise the single-use hose assembly by weakening the braid reinforcement commonly used in braid-reinforced silicone hose. This weakening and eventual failure will typically happen at the point of the retainment, to the fitting, to the hose.

To overcome this challenge where high-pressure requirements were needed, it was standard practice to use non-single-use hose assemblies with stainless-steel fittings and collars, which would then be subject to the requirements of cleaning and sterilization between operations. There is a market need for a single-use high pressure hose assembly that is able to withstand the pressure fluctuations of chromatography and filtration.

Avantor, having expertise in single-use manufacturing, including areas such as hose fitting and retainment design, recognized this common failure point and designed a new high-pressure fitting (SterilEnz). The new fitting uses a multi-barb design and, when used in conjunction with the proper retainment system, offers a large surface area, multi-point attachment to address high-pressure fluctuations common in downstream applications. Proper fitting and retainment design ensures that the connection between fitting and hose can withstand pressure fluctuations and the integrity of the system.

 Can you describe the general schematic of a single-use-technology-based hose assembly; does this include components such as valves, connectors, clamps, etc.?

 In general, the operation has two points, one inlet and one outlet, unless this is a manifold assembly, in which case there could be multiples of those inlets. These fluid pathways can be controlled using external valves and use connectors to change fluid direction. Many times, these pathways can be included in process instrumentation to measure process parameters, such as pressure and temperature.

 What impact do downstream single-use systems have on titer/yield?

 Any fluid pathway, when designed poorly, can develop hold-up volume that can negatively affect the titer or yield in a downstream process. Good engineering practices and knowledge of single-use systems will ensure proper design of these fluid pathways, preventing line losses and improving the yields in downstream processing.

Another area of focus that can have an impact on titer/yield in downstream is buffer preparation. Buffers play a significant role in biopharmaceutical manufacturing, typically representing the major component by volume used in biopharma downstream processing steps to help control pH levels and stabilize reactions during purification.

Traditional buffer preparation in monoclonal antibody (mAb) manufacturing requires large bioreactor tanks to mix the buffers from raw materials, such as salts (e.g., sodium chloride) in concentrated form that are then diluted with water-for-injection (WFI). Buffer preparation tanks not only require a large footprint but are also a significant capital investment. Every buffer produced must also be tested rigorously, requiring time and effort from a manufacturer’s valuable quality control and assurance resources. Each buffer must then be stored in a dedicated space or container, where the quality of the buffer has to be maintained until it is used, further adding to the facility space requirements.

An alternative solution is for a manufacturer to obtain ready-to-use buffers at 1x concentrations, which can also be packaged in single-use bags. Equipment options, such as in-line dilution systems, can help further improve a facility’s buffer management practices by offering the ability to prepare buffers on-demand. This allows a manufacturer to directly use the buffers in the process, eliminating the time it would normally take to prepare, mix, and store buffers in large tanks in its facility.


Vol. 34, No. 4
April 2021
Pages: 24–25

When referring to this article, please cite it as F. Mirasol, “Fluid Handling Systems are Seeing Benefit from Single-Use Technologies,” 

Downstream Processing

The adoption of single-use technologies in fluid-handling systems helps streamline downstream bioprocessing operations.

Fluid Handling Systems are Seeing Benefit from Single-Use Technologies

The past year has been a busy one for vendors and contract development and manufacturing organizations (CDMOs) that provide lyophilization equipment and services. On the bulk lyophilization side, demand for lyophilization for antibody-drug conjugates (ADCs), monoclonal antibody therapies (mAbs) for COVID-19, and some of the raw materials used in COVID-19 vaccines has been growing, says Scott Ross, global product specialist for bulk lyophilization with W.L. Gore and Associates.The company’s Lyoguard trays are being used in the production of mAbs; diagnostics tests for COVID-19 virus or antibodies; and some of the key raw materials used in messenger RNA (mRNA) vaccines, including synthetic oligonucleotides, adjuvants, and lipid nanoparticles, he says.

“For ADCs, lyophilization is likely to become a de facto standard,” Ross says, noting that operator safety concerns and the need to reduce turnaround times at CDMOs are driving use of contained tray systems. In addition, real-time monitoring of temperature and other parameters is becoming routine, he says.

As it has lifted demand for bulk lyophilization, the COVID-19 pandemic has also highlighted deficiencies with vial-based lyophilization, a process that has seen few changes in the past 50 years. By offering a powdered form of vaccine that could be shipped at ambient temperatures and reconstituted at the point of administration, lyophilization could have simplified the distribution of new vaccines, including those based on mRNA, which must be transported and stored as frozen liquids at extremely low temperatures, then mixed and manipulated before patients can be injected. Of the five COVID-19 vaccines that are now being used globally to prevent COVID-19, only Russia’s Sputnik V—which also comes in frozen liquid form—actually uses traditional vial freeze drying.

In the end, it’s a question of capacity. It would take three years for the industry to have enough capacity to lyophilize 300 million vials of product, says Alina Alexeenko, professor of chemical engineering, aeronautics, and astronautics at Purdue University and cochair of LyoHUB, a consortium of users and equipment and services vendors which was established four years ago to promote best practices and establish a roadmap for the future of this technology.

Fellow co-chair and Purdue professor Elizabeth Topp, who is also chief science officer at the Dublin, Ireland-based National Institute of Biopharmaceutical Research and Training (NIBRT), agrees.“Global lyophilization capacity wouldn’t allow us to process a billion vaccines a year, and stability testing is another hurdle. If you’re pushing a vaccine out the door in nine months for emergency use authorization, you clearly won’t have time to do a shelf stability study on a lyophilized formulation and guarantee 18 months stability,” she says. However, new analytical methods and process development strategies promise to improve processing and simplify distribution, administration, and the cold chain. “We can expect to see more lyophilized vaccines in the future,” says Steve Nail, senior LyoHUB scientific advisor who was formerly a Purdue professor and is now principal scientist at Baxter Biopharmaceutical Solutions, a CDMO in Bloomington, Ind.However, he says, the equipment used to process the materials will look quite different from the traditional stainless steel lyophilizers, with heat-transfer fluid circulating through shelves.

“The technology is up for disruption,” says Alexeenko. She notes that innovative drying processes (e.g., spray-freeze drying [both continuous and non-continuous]; spin freezing; thin- film freezing; and microwave drying), show promise to simplify the processing, transportation, and distribution of biopharmaceuticals and vaccines.

Lyophilization poses challenges, particularly for some proteins. A number of stresses are associated with freezing and freeze drying, as well as with agitation and phase interfaces, says Nail.Another problem is the fact that lyophilization involves very slow cooling, at a rate of one degree Kelvin per second, says Bill Williams, a professor at the University of Texas and inventor of the thin-film freezing process, which he developed years ago at the Dow Chemical Co.

TFF Pharma licensed his technology and commercialized it in 2019. Williams and his team, with corporate and US government funding, are now focusing on research designed to optimize use of thin-film freezing to improve the processing and delivery of biologics, including vaccines, along with the cold chain. Some of the problems that can occur with lyophilization are due to ingredients within vaccine formulations. As Williams notes, 80% of all existing vaccines use aluminum hydroxide adjuvants. If these ingredients freeze and then melt, particles can then aggregate, affecting both vaccine immunogenicity and activity, he says.

However, Williams notes, slow freezing is another challenge with lyophilization that affects vaccine activity. “When the freezing rate is 100 to 1000 times faster than it is for traditional lyophilization, as it is in thin-film freezing, the integrity of protein materials is maintained,” he says. Using thin-film freezing, the vaccine can either be reconstituted at the point of use, or administered via nasal or pulmonary delivery, he says.

Currently, Williams’ lab is working with TFF Pharma and more than 25 different pharma companies on research that is formulating liquid mAbs,mRNA lipid nanoparticles, and other bioformulations into thin-film powders that can be delivered to the lungs. The company recently worked on a reformulation of remdesivir as an inhaled dry powder, and in July 2020, completed Phase I of a clinical study that studied formulation of voriconazole as an inhaled powder.

Protein destabilization is another issue with lyophilization. Although research has studied the mechanics of chemical destabilization, which occurs as the result of reactions such as deamidation, oxidation, and chain cleavage or “clipping,” the science behind physical destabilization processes such as aggregation is not yet fully understood, says Topp.

“Aggregation is hard to control because it is part of the protein folding problem. We don’t really understand how proteins fold, and we don’t understand how they aggregate either,” she says.


Solid-state hydrogen deuterium exchange (HDX), an analytical method that is being used in research at Purdue and NIBRT, allows proteins to be studied in the solid state at high resolution, and is improving understanding of protein stability, says Topp.“A lot of what seems to drive the performance of these materials has to do with the hydrogen bonds that help make up the structure of the proteins, and the hydrogen bonds between the protein and the matrix that surrounds it. HDX interrogates the hydrogen bond network, and, so far, results suggest that the better the network, the more stable the material,” she says.

Evidence suggests that aggregation may be driven by partial protein unfolding. In the future, Topp says, this research might yield a metric that would reveal how well folded each individual protein is, which could help determine whether a particular biopharmaceutical formulation will perform well or not, whether it is made bylyophilization or another process.

In addition, Topp notes, research suggests that measurements made with HDX correlate closely with the stability of the protein in storage. “If we can characterize these materials and the environment that the protein sees with sufficient resolution, then maybe we can start to replace time consuming 18-month to two-year stability studies with advanced analytical characterization,” she says.

This approach would likely start with comparability protocols and then move to other parts of regulatory filings. “Stability studies impede all kinds of process changes and improvements. We hope that we can put tools in place to change that,” Topp says.

Breakthroughs in analytics and alternatives to traditional freeze drying promise to reshape biological development and the cold chain.

BioPharm International-04-01-2021

Maximum Output Starts with Optimized Upstream Processing

Using Anion Exchange Membrane Adsorbers to Ensure Effective Virus Clearance of Challenging Parvoviruses

Taking a Breath: Advances in Inhaled Biologics

Single-Use Bags Help Maintain Aseptic Ingredient Transfer to Bioreactors

Fluid Handling Systems are Seeing Benefit from Single-Use Technologies

Beyond Lyophilization

Stability Testing of Protein Therapeutics Using DLS

Supply Chain Challenges Creating Hurdles to COVID-19 Vaccine Production

Pressure Mounts for FDA to Ramp Up Drug Inspections

Avoiding Unforced Errors in the Bio/Pharma Industry