Aligning mRNA Progress with Regulatory Streamlining

BioPharm Peer Exchange

Technologies to develop and manufacture biopharmaceuticals continue evolving

Streamlining Bioprocesses with Automation

Technological advances are aiding manufacturers to streamline aseptic processes.

Speeding Up Aseptic Processes

Quality Considerations for Using AI in Bio/Pharma

Ask the Expert:

Sterile filtration and use of sterile single-use materials provide the best solutions depending on vector size.

Advances in Sterilization Technologies for Overcoming Viral-Vector Manufacturing Challenges

Patrick Lavery is an Editor for the MJH Life Sciences brands Pharmaceutical Technology and BioPharm International, and their respective websites, 

. He previously spent nearly a decade as a news anchor, reporter, and producer at New Jersey 101.5 FM.

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The California-based contract development and manufacturing organization (CDMO) SK pharmteco, which focuses on small molecules and advanced therapies, announced on Oct. 2, 2024 that it was entering into a multi-year agreement with the German biotechnology company AaviGen to be the preferred manufacturing partner for AaviGen’s lead gene therapy product, AVG-101 (1).

According to a press release from SK pharmteco, AVG-101 is intended for the treatment of heart failure; AaviGen’s business specializes in the preclinical stage, and the company is a developer of curative gene therapies for both cardiovascular and cardiopulmonary diseases (1).

"SK pharmteco stands out from the global pool of suppliers because of its extensive experience in process development and manufacturing of adeno-associated viral-based gene therapies,” Martin Busch, PhD, COO of AaviGen, said in the press release. “Additionally, their recent investments in expanding manufacturing capacity provide us with exceptional support from the earliest stages of development to the commercial phase, especially for common cardiac disorders, such as chronic heart failure."

“We are thrilled to partner with AaviGen on their innovative gene therapy for heart failure,” added Patrick Mahieux, SK pharmteco president of cell and gene, Europe. “Our deep expertise in viral vector manufacturing and our commitment to quality and efficiency make us the ideal partner to support the development and commercialization of this promising therapy. We look forward to contributing to the advancement of this groundbreaking treatment for patients in need.”

In the press release, AaviGen’s proprietary adeno-associated viral vector technology was described as being able to enable development of therapies to target, and reverse, underlying molecular causes of heart muscle dysfunction, by concentrating on areas such as cardioprotection, contractility, and energy metabolism (1).

AaviGen has been around just since 2019, while SK pharmteco is a subsidiary of SK Inc., the strategic investment company of SK Group, which the press release identified as the second-largest conglomerate in South Korea (1).

"SK pharmteco is a key partner for AaviGen as part of our strategy to develop and commercialize innovative gene therapies against diseases of the cardiovascular and cardiovascular system,” said Patrick Most, AaviGen CEO, in the press release. “The SK pharmteco team offered the trust, competence, and understanding of how a young company like AaviGen must operate in a dynamic market from the first minute of our interactions. We look forward to the collaboration with great confidence."

In late September 2024, SK pharmteco announced another partnership, that with the supply-chain service company, Cryoport (2). The two businesses will work together to provide biopharmaceutical companies with integrated logistics and manufacturing services—such as end-to-end cell and gene therapy services that include biomaterial collection, bioservices, data solutions, and shipping systems.

1. SK pharmteco. SK pharmteco and AaviGen Announce Strategic Partnership to Manufacture AaviGen’s Lead Gene Therapy Product for Heart Failure. Press Release. Oct. 2, 2024.
2. SK pharmteco. SK pharmteco and Cryoport Partner for Cell and Gene Therapy Logistics and BioServices. Press Release. Sept. 25, 2024.

Articles

SK pharmteco will be the preferred manufacturing partner for AVG-101, AaviGen’s lead gene therapy product.

AaviGen, SK pharmteco Announce Manufacturing Agreement for Heart Failure Gene Therapy

Feliza Mirasol is the science editor for 

Evonik announced on Oct. 9, 2024 that it won the 2024 CPHI Excellence in Pharma Award in the “Sustainability” category for its plant-based squalene, PhytoSquene, an adjuvant that can be used for parenteral drug delivery applications. The product is the first plant-based squalene on the market that complies with good manufacturing practices (GMP), according to a company press release, .

Vaccines such as the H1N1 influenza vaccine use squalene as an adjuvant. Adding it to vaccines stimulates the body’s immune response to the active ingredient. Pharmaceutical-grade squalene is currently sourced from shark liver oil, however, which runs the risk of depleting shark populations and negatively impacting marine biodiversity, Evonik stated in its press release. “In terms of quality, animal-derived squalene may be susceptible to batch-to-batch inconsistencies which can complicate regulatory processes. Some patients also cannot use animal-based products for cultural or religious reasons,” the company also stated in the release.

PhytoSquene belongs to Evonik’s Next Generation Solutions (NGS) portfolio, in which products are categorized by a distinct sustainability profile. Evonik’s Health Care business unit aims to transform more than 70% of its portfolio by revenue to NGS by 2030. This goal also includes commitments from the company’s life sciences division, Nutrition & Care, to reduce the need for animal-derived products while growing its biosolutions portfolio, according to the company.

“Sustainability drives us! This award confirms that we are pursuing the right goals: to offer our customers alternatives that enhance patient outcomes and protect our planet,” said Yann d'Hervé, head of the Health Care business line at Evonik, in the press release.

PhytoSquene is manufactured in Germany using a proprietary process. It comes from amaranth (

) oil. Amaranth is an herbaceous plant cultivated in many parts of the world, making it a renewable feedstock. The manufacturing process produces consistent quality and is compliant with 

 regulations. Evonik launched commercial-scale PhytoSquene in 2023.

In 2022, Evonik won a CPHI Pharma Award in the category “Packaging and Drug Delivery” for a polymer-based messenger RNA (mRNA) drug delivery system. The drug delivery system is also used for delivery of other nucleic acid-based therapeutics.

Evonik also announced that it is transforming its Health Care business to focus on strategic core growth areas. Under the restructuring, Evonik will evaluate strategic options, such as partnerships or divestment, to develop its keto and pharma amino acid production sites in Ham, France, and Wuming, China. The company also plans to discontinue its production of keto acids in Hanau, Germany by the end of 2025 (1).

Evonik’s Nutrition & Care, the Health Care business within the company’s life sciences division, is transforming into a system solutions provider and is growing in core areas of expertise, including complex and highly potent APIs, oral and injectable drug delivery, and precision biosolutions, such as lipids for mRNA and gene delivery, and cell culture ingredients. The company described system solutions as tailored offerings that integrate products, technologies, and services with sustainability benefits, according to an Oct. 11, 2024 company press release (1).

For the full list of the 2024 CPHI Excellence in Pharma Awards winners, read the 

1. Evonik. Evonik to Restructure Its Keto and Pharma Amino Acid Business. Press Release. Oct. 11, 2024.

Evonik's plant-based squalene, PhytoSquene, used in parenteral drug delivery applications, won the CPHI Excellence in Pharma Award in the “Sustainability” category. 

CPHI Award Granted to Evonik for Plant-based Adjuvant Used in Parenteral Drug Delivery

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The European Medicines Agency (EMA) and the Heads of Medicines Agency (HMA) announced that the draft joint strategy for the European Union to 2028 (EMANS 2028) is now open for public consultation. The consultation period will be open until Nov. 30, 2024.

 has been reviewed and updated as regulatory and technical situations have changed rapidly (1). New regulations (e.g., legislation to address public health emergencies and reform of the EU medicines regulations) and technical advances, such as the use of artificial intelligence (AI), have spurred the changes to the original strategy.

“Since EMANS 2025, the availability and use of AI have become more commonplace, and the network aims to harness the full potential of AI across all the EMANS focus areas. The use of digital technology and AI has the potential to transform the network’s core work of evaluating human and veterinary medicines for authorization and monitoring their safety through the EU’s pharmacovigilance system. For veterinary medicines, the network will also push ahead with its vision to implement new digital solutions in its European Veterinary Big Data strategy,” the strategy document states (1).

The strategy to 2028 will focus on creating pathways to medicines in the EU. There is also a plan to find ways to leverage data, digitalization, and AI to improve processes.Innovation and competitiveness are priorities that will be nurtured through a regulatory science and research environment, according to EMA. Strengthening the supply chain and the sustainability of the EU network are also on the agenda. Each focus area will be evaluated by EMA and HMA to assess progress and adapt approaches, if necessary.

“The six focus areas have been carefully chosen to support the network’s core work of evaluating medicines as we take strides to promote the development of medicines and ensure that they reach those who need them,” said Maria Lamas, chair of the HMA Management Group, in a press release (2).

“The overarching theme of our updated strategy to 2028 is that of change—rapid, somewhat unpredictable, but nonetheless full of promise,” said Emer Cooke, EMA executive director, in the press release (2). “It will guide the network as it seizes opportunities and meets the challenges of the near future, including preparing for and responding to public health emergencies and threats such as antimicrobial resistance. Now is the time to share your views and help shape the direction of the network for the next few years.”

Stakeholders may provide input through an online questionnaire until Nov. 30, 2024. [https://ec.europa.eu/eusurvey/runner/EMANS2028publicconsultation] The strategy will be finalized after the consultation period and be implemented through multi-annual workplans. The final strategy is expected to be adopted by March 2025.

“While EMANS 2028 cannot preempt the legislative changes or predict the future, this strategy will prepare the network for the revisions to the pharmaceutical legislation and further changes to the technological and regulatory landscape,” Cooke says in the document’s introduction.

Seizing Opportunities in a Changing Medicines Landscape, The European Medicines Agency Network Strategy 2028 (draft)

EMA. Seizing Opportunities in a Changing Medicines Landscape. Press Release. Oct. 9, 2024. 

The draft joint network strategy for the EU until 2028 will be open for public consultation until Nov. 30, 2024.

EMA and HMA Publish Network Strategy for Public Consultation

Biologics and other complex biomolecule modalities present challenges to fill/finish steps. Being able to ensure a sterile environment during aseptic processing is one of them.

The aseptic/fill finish process for biologics can be challenging because of the sensitivity of biologic products. In addition, where the product is located in the manufacturing process at a given time makes aseptic fill/finish difficult, says Michael Francis, senior product manager, CPC Biopharma.

According to Francis, two primary challenges are maintaining sterility and minimizing contaminants, and, because the fill/finish steps for complex modalities such as cell therapies, for example, are the last processing steps before a product is delivered to a patient, biomanufacturers must carefully consider patient safety along with the high costs—in time and money—if a batch is lost and has to be reproduced.

“Biologics are often more sensitive to contamination with bacteria, viruses, or particulates, so maintaining sterility is paramount. Even with advanced automation, human intervention can still be the source of contamination risks, so ensuring operators can easily and stringently adhere to aseptic techniques is critical,” Francis states.

In addition, one of the most critical responsibilities in biopharma aseptic fill/finish is ensuring the integrity and viability of the molecule, explains Jackie Klecker, executive vice-president and general manager at Lifecore Biomedical, a US-based contract development and manufacturing organization. Ensuring molecular integrity and viability requires careful and accurate characterization of the key properties of these highly complex, sensitive, and expensive products. “Each molecule differs from the next and any number of forces can negatively impact the product, including light, temperature, shear, oxygen, and product contact, among others,” Klecker says.

Pirkko Kortteinen, PhD, chief operating officer, 3PBIOVIAN, agrees that product stability is one of the most significant challenges for the aseptic fill/finish of biologics and complex biotherapeutics. “The stability of large complex molecules such as proteins, living virus particles, or viable cells can be compromised during aseptic fill/finish by environmental factors such as temperature, high pressure, metal ions, adsorption agitation, or shear stress,” she adds.

“The aseptic fill/finish process must be designed to preserve molecule structures, biological activity, and cell viability and avoid degradation, aggregation, or other damage. Many products cannot withstand holding at room temperature,” Kortteinen further emphasizes.

“Understanding the limitations of each molecule is essential to establishing and maintaining expert control of the optimal environment, across all aspects and timepoints of the fill/finish process,” says Klecker. “This can include strict temperature control requirements/timing during production steps, such as controlled thawing timing, as well as use of low-shear peristatic pumps for filling.”

Meanwhile, Kortteinen explains that, due to the nature of these biopharma products, total processing time must be kept short—in hours rather than days, in some cases—to complete the operations. Therefore, all processing steps, including melting, sterile filtration (if applicable), filling, closing, visual inspection, sample withdrawal, and secondary packaging must be carried out within a tight time frame.

The sensitive nature of biotherapeutics limits choices of sterilization methods, so, although heat sterilization in the final container is recommended, it is most often impossible, Kortteinen points out. Sterile filtration of the final bulk solution for many biological products must instead be performed before aseptic fill/finish. In contrast, a final cell therapy product, which consists of viable cells or cell-derived matrices that are not amenable to final sterilization or filtration, may instead require aseptic techniques throughout the manufacturing process, Kortteinen remarks.

Establishing an aseptic fill/finish strategy

Francis also notes that, because many aseptic fill/finish operations are performed in an isolator, aseptic fill/finish for biopharmaceuticals can be extremely expensive, and the equipment challenging to use. “Open manipulations within the isolator require good aseptic handling procedures to be successful. Incorporating single-use systems (SUS) into these applications has helped reduce contamination and particulate risks; however, these SUS designs are often very complex, sometimes resulting in ‘tubing spaghetti’,” Francis states. “These overly complex systems can be very difficult for an operator to handle—and any difficulty in assembly can introduce unintended risks of operator error.”

Furthermore, minimizing product loss at the aseptic fill/finish stage is critical because the starting materials, and therefore the drug substance, for biotherapeutic products are extremely valuable, Kortteinen points out. “Minimizing the loss of volume during processing is an important target for fill process designers,” she states.

“Facilities with the flexibility to implement creative solutions, whether it be utilizing state-of-the-art equipment with advanced control features or optimizing aspects of the filling process, are best equipped to minimize product waste,” Klecker adds.

Meanwhile, product freezing and shipment, which are more common with biotherapeutic products, also present challenges, Francis explains. Kortteinen points out that many biotherapeutics, especially in their early product life cycle, are stored at low temperatures, such as -80 °C or cryopreserved in liquid nitrogen. These low storage temperatures challenge container integrity during cryopreservation, she specifies.

“System designers need to consider component materials that can handle the wide temperature ranges associated with cryopreservation and subsequent thaw cycles that many biologic products undergo,” Francis adds. “Component suppliers should be able to provide leak, burst, microbial ingress, and other testing that demonstrates how their products perform throughout the entire process and associated activities [such as] sterilization, shipping, and freeze-thaw.”

Overall, notes Klecker, the complexities and specific requirements of biologics manufacturing necessitate a high level of expertise and meticulous process management to address these challenges effectively.

Solving the problems encountered during aseptic fill/finish of biopharmaceuticals was greatly helped by the advent of single-use technology (SUT).

SUTs have seen vast development in the past two decades, emphasizes Kortteinen, who explains that single-use supplies, typically sterilized plastic components or assemblies, have changed aseptic fill/finishing processes by adding operational efficiencies.

“Using single-use filling lines for fluid transfer can shorten overall manufacturing time,” Kortteinen says. “By using ready-made and sterilized assemblies, one can avoid time-consuming and laborious campaign changeover procedures of equipment. The need for sterilization in place for the fill line disappears when the suppliers sterilize the disposable fill line. Also, using high-quality, high-capacity disposable sterilizing grade filters shortens the manufacturing time.”

A significant benefit to using single-use materials in aseptic fill/finish is the minimization of cross-contamination risks, which is a crucial concern, especially for multi-product facilities and contract manufacturing organizations, Kortteinen remarks.

However, Francis explains that, although SUTs such as disposable filling lines and single-use bags help reduce cross-contamination risks and simplify cleaning and validation processes, manufacturers still need to ensure that the SUS components perform as intended within their processing parameters. “For example,” Francis says, “newer polymer SUS must be shown to withstand the applicable processing temperature and pressure levels that traditionally posed no issues for stainless-steel equipment.”

“Previously,” explains Kortteinen, “fill/finish manufacturers washed and sterilized primary packaging materials in-house. The processes needed heavy equipment and facility investments. Validation of such quality critical processes and qualification and maintenance of equipment and systems was labor-intensive.” In comparison, today, high-quality ready-to-process vials, stoppers, and other packages are available from suppliers.

“Outsourcing the preparation and sterilization of primary packages has simplified the fill and finishing operations. Validation activities when using ready-to-process components focus on material transfer and loading sterile components into the filling line to demonstrate that all components are kept sterile throughout the processing,” Kortteinen states.

Francis further notes that, to create optimal systems, manufacturers should ask component suppliers for key information on chemical compatibility, flow performance, life cycle testing, extractable data, and product integrity in freeze-thaw cycles, among other information. “Many new therapies are being cryopreserved at temperatures beyond -150 °C, for example, so the components in the fill/finish stage need to deal with these ultralow temperatures,” he says.

Relying on the latest disposable systems, utilizing low shear peristatic pumps within fillers, and streamlining the isolator filling line setup to maximize yield and minimize opportunities for contamination are also important solutions, says Klecker. “Equally important as these in-process solutions are the extensive testing protocols that we develop with our customers to incorporate into the fill/finish process. These play a key role in not only characterizing the molecule and identifying its vulnerability to external factors, but also in monitoring for any unwanted impact during the aseptic fill/finish work,” she adds.

Klecker also emphasizes the point of “keeping the end in mind” for each product’s commercial fill/finish process. “By working to learn about and address any unique, product-specific behaviors early in our work with a molecule,” she stresses, “we can ensure that our final process is optimized for the valuable commercial lots we will produce.”

Manufacturers these days are also looking for ways to simplify SUS that make their way into the isolator, according to Francis. For instance, sterile connectors allow SUS to be used outside of the isolator, where the connectors function as a passthrough on the beta port. “Removing the surge bag and even a portion of the tubing manifold, for example, can reduce the ‘tubing spaghetti’ that otherwise complicates handling by an operator. And as part of a closed system, aseptic connectors help stop particulates and contamination from entering the flow path when connecting filling equipment,” he explains. “These connectors also make modular systems possible by connecting components within the systems such as tubing, bags, filters, and manifolds. The integration of closed processing and modular systems enables efficient, flexible workflows while providing the aseptic processing biologics manufacturers need to minimize contamination risks.”

Filling line manufacturers have already developed innovative new designs and technologies for aseptic fill/finish that are suitable for biotherapeutics, points out Kortteinen. For instance, small batch sizes used in gene therapy and cell therapy have created a need for small automatic filling lines suitable for multi-product fill/finish operations in cell and gene therapy manufacturing facilities. Meanwhile, automatic filling lines improve quality by avoiding human interventions, and automation adds operational efficiencies and enables effective scaling up of fill/finish operations, she notes.

“[T]he industry will continue to optimize facilities and equipment to improve the ability to monitor for and address external factors that may jeopardize the viability of complex biotherapeutic products,” remarks Klecker. These innovations may include: the addition of continuous temperature controls throughout the entire fill/finish process and more low-temperature storage capacity; greater adoption of disposable components to reduce burden of cleaning and cleaning validation; and facilities that are designed with specific light spectrum ranges.

Meanwhile, Francis expects that there will continue to be greater use of closed SUS, which he points out are adaptable to any scale or system depending on a unique biologic or therapy needed. “SUS allow smaller, modular segments to process specific batches of product, so modularity will be more broadly deployed. Modularity helps simplify operator SOPs [standard operating procedures] and workflow, too, because all personnel use the same or similar systems.,” he says.

Francis also notes that the ability to continuously filter in upstream processing via perfusion is starting to create a continuous flow of product to the filling stage. This continuous processing, along with modular SUTs, is creating a future of faster manufacturing. However, while there is a need for more automation, Francis points out that increased automation as a solution may be difficult to implement because there is no single solution. “With more process data collection and quick analytics through AI [artificial intelligence], there’s an expectation that future filling will be tailored and precise. Automation can provide greater reliability, faster processes, and less risk of operator error, but current automation designs are often very complex so you can lose some of the benefits automation is intended to deliver. Also, there’s a need to standardize on those designs. The industry recognizes the benefits of standardization, but it’s tough to implement for a variety of reasons,” he explains.

Furthermore, Francis anticipates that biologics manufacturers will seek more efficient processes earlier in drug development to allow for more streamlined scaling because thinking ahead to the commercial manufacturing process while still in the drug discovery and efficacy testing stages saves time later on. “For example, the more manual and operator-dependent aseptic techniques such as tube welding and the use of biosafety cabinets might work fine in small-scale processes, but that doesn’t translate well when it comes to commercialization,” he says.

“Finally,” Francis adds, “many hurdles and challenges occur when researchers and developers, especially universities and start-ups in the [cell and gene therapy] space, look to transfer their technology or product from lab scale to commercial scale. As start-ups gain a broader understanding of how their products might eventually be produced—and if they begin incorporating those approaches into their own early-stage processes—the more seamless tech transfer can be.”

Kortteinen also highlights newer regulatory considerations that may affect the future of aseptic fill/finish. She brings up that the new Annex 1 guideline (1) introduced in Europe, for example, has a broader scope to include new technologies (e.g., isolators and restricted access barrier systems [RABS]), especially for new facilities or installations. Kortteinen explains that isolators can be completely closed and decontaminated with hydrogen peroxide, which minimizes contamination or cross-contamination risks. “Automatic filling in an isolator is a choice for viral products that require containment at biosafety level 2, as it protects operators from exposure to the products. In the future, aseptic [fill/finish] in either isolator or RABS will be an expectation,” she states.

“Annex 1 requires a unidirectional flow for the transfer of materials, equipment, and components. Separate change rooms for entering and leaving the filling are desirable,” Kortteinen summarizes. “These requirements will influence the cleanroom design of new facilities and may induce facility renewals in existing cleanroom setups.”

1. EC. Annex 1: Manufacture of Sterile Medicinal Products. 

®
Vol. 37, No. 9
October 2024
Pages: 8–11

When referring to this article, please cite it as Mirasol, F. Understanding Molecule Sensitivity in Aseptic Fill/Finish. 

Aseptic fill/finish of biopharmaceuticals requires an understanding of the structure and limitations of each molecule.

Understanding Molecule Sensitivity in Aseptic Fill/Finish

Dayna Turner is group leader, Science & Technology, at STERIS, Saint Louis, Mo.

Dijana Hadziselimovic is laboratory manager, Technical Services, at STERIS, Saint Louis, Mo.

Paul Lopolito is a technical services specialist at STERIS, Mentor, OH.

Mary Schanne is senior account manager at STERIS, Saint Louis, Mo.

Amy Thanavaro is manager, Science & Technology, at STERIS, Saint Louis, Mo.

Jeff Felker is deputy director, Cleaning Validation SME, Sanofi MSAT Swiftwater, Pa.

A standard pharmaceutical or biopharmaceutical process equipment cleaning cycle consists of an initial rinse, caustic wash, intermediate rinse, acid wash, and then a series of post-acid rinses ending with a final purified water rinse and drying step. The current good manufacturing practice (CGMP) regulations define critical process parameters (CPPs) and predetermined critical quality attributes (CQAs) applicable to a cleaning cycle. The desire for a more environmentally sustainable cleaning cycle had previously led to recovery of final rinse water to be used for the initial rinse step; change to formulated chemistries to reduce chemical consumption, impact on the environment and waste treatment; and change in cleaning temperature to decrease energy and cycle time. In this article, the authors explore removing the intermediate rinse and blow down step before the acid wash for a biopharmaceutical upstream and downstream process step resulting in both rinse volume and time savings. A short blow down will still be recommended to reduce residual liquid in the system between washes.

An acid wash may not be required for all cleaning cycles, but this step helps remove inorganic residues (e.g., iron oxide) on the surface, provides continuous maintenance of the chromium oxide passive surface, neutralizes wastewater, and reduces bioburden.

The elimination of a water rinse and blow down between the caustic and acid wash steps seems practical, but before being considered, an evaluation should be performed with an assessment of the risks, controls in place to mitigate the risk, and monitoring of the cleaning process to ensure continuous control of the validated state. A list of questions and answers (see Table I) based on the International Conference for Harmonisation (ICH) Q9 

 guideline helps in the risk assessment process (1). Common sources of hazards in the pharmaceutical industry include handling and storage, human error, emissions, use of volatile organic compounds, reactions, and leaks of effluents (2). The identified concerns from the risk assessment include: a change in the cleaning solution temperature due to an exothermic reaction when mixing caustic and acid solutions, potential off gassing of chemical compounds, and redeposition of residues removed during the caustic wash step (3–5). To investigate these concerns, it is important to know the exact chemical formulation of both caustic and acid cleaning agents. The physical properties of the acid rinse solution were assessed with temperature and pH checks, using different ratios of caustic and acid solutions. The volatile chemicals were assessed using gas chromatography with mass spectrometry (GC–MS). The cleaning and rinsing of the caustic cleaner, followed by the acid cleaner was also investigated using 304 stainless-steel coated surfaces.

Testing strategy to investigate process risk

All changes to CGMP processes require a change review, and based on the significance of the change, a new validation or revalidation may be required. For an automated cleaning process, this may include repeating the spray device coverage testing, ensuring the cleaning equipment has been commissioned and validated, and confirming that the rinse water recovery is consistent with surface cleanliness.

Submitted: Aug. 5, 2024
Accepted: Aug. 30, 2024.

, Step 4 version (2005).
2. Bhusnure, O. G.; Dongare, R. B.; Gholve, P. S.; et.al., Chemical Hazards and Safety Management in Pharmaceutical Industry. 

 (3).
3. Dermaut, W. Chapter 3: Process Safety and Reaction Hazard Assessment. In 

Chemical Engineering in the Pharmaceutical Industry: Active Pharmaceutical Ingredients

,2nd ed.; am Ende, D. J.; am Ende M. T., Eds.; John Wiley & Sons, 2019. DOI:10.1002/9781119600800
4. Allian, A. D.; Shah, N. P.; Ferretti, A. C.; et.al., Process Safety in the Pharmaceutical Industry—Part 1: Thermal and Reaction Hazard Evaluation Processes and Techniques. 

 (11), 2529–2548. DOI: 10.1021/acs.oprd.0c00226
5. Agarwal, P.; Goyal, A.; Rajat, V. Chemical Hazards in Pharmaceutical Industry: An Overview. 

 (2). DOI: 10.22159/ajpcr.2018.v11i2.23160

Dayna Turner is group leader, Science & Technology; Dijana Hadziselimovic is laboratory manager, Technical Services; Paul Lopolito is director, Technical Services; Mary Schanne is senior account manager; and Amy Thanavaro is manager, Science & Technology; all at STERIS, Saint Louis, Mo. Jeff Felker is deputy director, Cleaning Validation SME, Sanofi MSAT Swiftwater, Pa.

®
Vol. 37, No. 9
October 2024
Pages: 23–28

When referring to this article, please cite it as Turner, D.; Hadziselimovic, D.; Lopolito, P.; et al. Strategic Changes to a Legacy Cleaning Approach Result in a More Sustainable Process. 

In this article, the authors explored the elimination of a water rinse and blow down following the caustic wash step, examining potential safety considerations, the effect on the quality of the cleaning process, and the potential benefit of implementing this change.

Strategic Changes to a Legacy Cleaning Approach Result in a More Sustainable Process

At CPHI Milan held in Milan, Italy, on Oct. 8–10, 2024, CPHI announced the winners of the 2024 Excellence in Pharma Awards (1). The winners are:

Future Leader Award: Megan Smyth, Almac Sciences

Woman of the Year Award: Julia Tsetis, OFET Pharmaceutical Group

CEO of the Year: Guy Clark, CNX Therapeutics

API Development and Innovation: Lonza, AI-Enabled Route Scouting Service

Drug Delivery and Device Innovation: Adare Pharma Solutions

Finished Formulation: Unistraw, UniLayer Straw Delivery System

Manufacturing Excellence: RheaVita, RheaLyo GMP-Flex continuous freeze-dryer

Packaging and Machinery: Smurfit WestRock, CereScreen Digital Display Label

Regulatory and Compliance: ProPharma Group, Prodigy

Start-Up Initiative: Acu-Flo Ltd, Nebu-Flow nebulizer platform

Sustainability Award: Evonik Industries AG, PhytoSquene

Heart of Pharma Award: West Pharmaceutical Services

Accelerating Innovation Award: AustinPx, KinetiSol Technology.

The new Future Leader category was awarded to Megan Smyth of Almac Sciences for rising through five levels in her seven years at the company and for being awarded a UK Research and Innovation (UKRI) Future Leaders Fellowship. “Megan has built an innovative flow chemistry platform at Almac and there is a clear strategic alignment between her career intentions and the future requirement for leadership within the company,” CPHI stated in the press release.

The other new category, Woman of the Year, went to Julia Tsetis, CEO of OFET Pharmaceutical Group, for her commitment to innovation, sustainability, and social responsibility. “In her role as CEO of the OFET Pharmaceutical Group she is regarded as a true visionary and trailblazer in the pharmaceutical industry and has left an indelible mark on the global healthcare landscape,” the press release stated.

Guy Clark, CEO of CNX Therapeutics, was awarded CEO of the Year for significantly growing the company and transitioning it to a standalone entity with £50 million in revenues with more than 20 products sold worldwide.

The API Development and Innovation category went to Lonza for its AI-Enabled Route Scouting Service, which uses artificial intelligence (AI)-enabled predictive tools “that combine theoretical retrosynthesis with supply chain intelligence to identify commercially viable routes. This crystallizes the company’s advanced, data-driven approach to retrosynthetic analysis for designing optimal synthesis routes for small molecule APIs.”

Adare Pharma Solutions won the Drug Delivery and Device Innovation category for its tremors and other uncontrolled body treatments. “Based on an innovative sprinkle formulation, it is a game-changer for patients who struggle to take capsules due to hand tremors or difficulty in swallowing as it can be easily mixed into soft foods or liquids,” CPHI stated.

The Finished Formulation award was given to Unistraw’s UniLayer Straw Delivery System 2mg Loperamide HCl straw, which delivers medication via a plant-sourced and fully compostable polymer straw. This sustainable solution enhances compliance and reduces misuse.

The RheaLyo GMP-Flex continuous freeze-dryer +1 2 from RheaVita was given the award in the Manufacturing Excellence category. It is the first continuous freeze-drying system for aseptic good manufacturing practice production.

CereScreen Digital Display Label from Smurfit WestRock received the Packaging and Machinery award for its seamless integration of digital screens into clinical trial investigational medicinal product packaging. “The screen allows for multiple pages and contains all required label content, including QR codes and other information. In addition, the labels can be customized to suit the specific needs of the trial, including support for multiple languages,” CPHI stated.

The Regulatory and Compliance category was given to Prodigy from ProPharma Group. The suite of AI and advanced automation applications solve productivity challenges that can cause delays and incur costs. “It effectively removes delays, de-risks program delivery, shortens time-to-insight and enhances the overall medicinal value of its clients’ assets,” the press release stated.

Acu-Flow Ltd’s Nebu-Flow nebulizer platform received the Start-Up Initiative Award for overcoming technical limitations by “generating a soft aerosol, with high inhalable fraction (>90%), which can aerosolize biological formulation, such as RNA.”

The Supply Chain Excellence category was awarded to QYOBO GmbH for “a platform that provides crucial insights into global supply chains to help companies identify, manage, and anticipate supply risks. With data from over 250 sources, QYOBO offers a comprehensive view of supply chains for over two million drugs in over 40 countries, enabling quick, informed actions to mitigate risks and prevent drug shortages.”

Evonik Industries AG received the Sustainability Award for their PhytoSquene plant-based squalene for parenteral applications. “This is the first plant-derived, GMP-quality squalene. Pharmaceutical-grade squalene is currently sourced from shark liver oil, which depletes shark populations and negatively impacts marine biodiversity, whereas PhytoSquene is made from amaranth oil from the herbaceous plant 

, which can be grown as a renewable feedstock in many regions of the world.”

The Heart of Pharma award went to West Pharmaceutical Services for its collaboration with the Fox Chase Cancer Center, a National Cancer Institute (NCI)-designated Comprehensive Cancer Center, in the delivery of cancer screenings to residents of Pennsylvania via a mobile screening unit. 

AustinPx received the Accelerating Innovation Award for its KinetiSol Technology, which is a fusion-based, solvent-free process designed to address poor drug solubility. “As new molecules emerge with complex properties, traditional formulation methods struggle to meet the demand for effective drug absorption. KinetiSol disrupts the status quo by rapidly converting crystalline drugs into amorphous solid dispersions, significantly enhancing bioavailability.”

“CPHI continues to drive the future of pharma by creating a platform where innovation thrives, and the brightest minds can exchange ideas and form impactful partnerships that elevate the entire pharmaceutical industry,” said Tara Dougal, Brand and Content Director at Informa Markets, in the press release. “I am thrilled to congratulate the winners of 2024 CPHI Pharma Awards, with special recognition to those honored in our new categories, the Future Leader Award and Start-up Initiative Award. These trailblazers are not just advancing drug discovery and delivery but inspiring the next wave of innovation and leadership in pharma. A heartfelt thank you to everyone who participated and to our esteemed jury, whose insights have helped spotlight the very best our industry has to offer.”

1. CPHI. CPHI Excellence in Pharma Award Winners 2024. Press Release. Oct. 8, 2024.
2. CPHI. Awards—CPHI Milan. 

cphi.com/en/europe/cphi-celebration/awards.html

This year's CPHI award winners emphasize achievements in pharma excellence.

CPHI Announces Excellence in Pharma Awards (CPHI Milan 2024)

Nicolas Camper, senior director—Bioconjugation Chemistry, Abzena, discussed new advancements in ADC technologies, including the role of complex chemistries.

Exploring New Advancements in ADC Technologies (CPHI Milan 2024)

Nicolas Camper, senior director—Bioconjugation Chemistry at Abzena sat with the Pharmaceutical Technology Group during CPHI Milan to discuss  advancements in conjugation and linker technologies in antibody-drug conjugate (ADC) development and manufacturing.  

Nicolas Camper, Senior Director—Bioconjugation Chemistry, Abzena. Image courtesy of Abzena.

“There have been a lot of new developments in terms of conjugation and linker technologies. There is a definitive move towards site-specific conjugation technologies and away from stochastic conjugation approaches that allows [us] to prepare better-defined antibiotic-[drug] conjugates,” Camper said.

Another new development that has been observed over the past few years is that linkers are becoming more sophisticated. “Quite often, when you conjugate cytotoxic drugs, they are quite hydrophobic, and that can impact the performance of your ADC. So, there has been a lot of effort put into developing more hydrophilic linkers,” he noted.

Finally, another noticeable advancement in this area has been the development of branch linkers. Branch linkers give some options to prepare ADCs and modulate the drug loading, Camper explained. “[T]hat is something relatively new and should have an impact on the performance of ADCs and ultimately result in better ADCs and more programs,” he stated.

In his role at Abzena, Camper has scientific oversight of the group’s R&D activities focusing on ADC design and developability for external clients. His responsibilities involve technical management of pre-clinical ADC development projects from discovery stage to lead candidate selection. The group has expertise in synthetic chemistry, assembling linker-payloads, conjugation of the linker-payload to various types of protein carriers (primarily antibodies), and analysis of these bioconjugates.

Attendees can visit Abzena at Booth 18H11. CPHI Milan is being held from Oct. 8–10, 2024 in Milan, Italy.

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At CPHI Milan, Nicolas Camper, senior director—Bioconjugation Chemistry, Abzena, spoke about new developments in ADC technologies, including the role of complex chemistries.

Biopharmaceutical manufacturing largely involves biomolecules whose stability depends on temperature. Antibodies, insulin, and, more recently, messenger RNA (mRNA)-based vaccines are examples of biotherapeutics that require cold storage. This article explores the difficulties with developing stability-enhancing formulations for temperature-sensitive biomolecules and technology innovations that are enabling their formulation.

Factors that have been most challenging in developing formulations that enhance stability in temperature-sensitive biomolecules include buffer, pH, and sugar. These are usually the typical factors that need to be evaluated to thermodynamically stabilize monoclonal antibodies (mAbs), notes Joseph Cao, PhD, senior scientist, Analytical and Formulations Development, Bionova Scientific. He explains that there are different types of buffers, sugars, and a range of pH to choose from when developing a proper formulation for mAbs that will prevent aggregation, degradation (such as deamidation, oxidation), and particle formation. Besides those factors, amino acids, salts for ionic strength, and excipients are also important in developing a formulation recipe, he adds.

In addition, the limited amount of therapeutic protein to work with poses another challenge to formulation development. “With design of experiment (DoE), it has allowed us to cut the amount of protein and help us efficiently screen for factors,” Cao says. “Freeze-thaw cycles, container combability, and manufacturing impurities should also be considered during formulation development.”

Heonchang Lim, director, Formulation Development, Samsung Biologics, agrees that finding the ideal range of pH and buffer conditions while selecting the optimal formulation condition to get a good thermal stability property are primary factors that challenge formulation development in these molecules. However, he notes, the thermal stability problem can mostly be solved by using optimal pH buffer and excipient combination.

“Nowadays, we bump into another challenge to develop an optimal formulation condition. That is regarding solubility and hydrophobicity of the molecule. Though clients would prefer higher concentrations, when taking unique characteristics of molecules into consideration, formulation scientists cannot always get high enough concentrations. In addition, pH shift may occur due to the Donnan effect,” Lim says. “Consequently, when a client requests a highly concentrated formulation, particularly above 100 mg/mL, [the] formulation team at Samsung Biologics implements rigorous gate checks to prevent such challenges from arising during formulation stages.”

Caroline Ailhas, Pharmaceutical Development director, CARBOGEN AMCIS says that, for her, the main factors challenging formulation development for temperature-sensitive molecules revolve around maintaining the structure and functionality of the molecule and defining the required duration of stabilization. The latter can vary from a few days—to allow for manufacturing and visual inspection before storage at 2–8 °C—to several months, or even years. “The definition of this duration significantly impacts the formulation strategy,” Ailhas says.

“One of the key performance indicators for the development of biotherapeutic formulations is colloidal stability to prevent protein aggregation and potentially precipitation,” Greg Chrimes, site head and vice-president, Analytical and Formulation Science, KBI Biopharma, adds. He points out that proteins may denature and aggregate at interfaces such as air/liquid, liquid/liquid, and liquid/solid. Chemical modifications and degradation may also impact effector function, efficacy, conformational stability, and other important parameters, he emphasizes.

“Excipients are evaluated during formulation development to limit product denaturation that may lead to particle formulation or chemical modification. Those may include buffering components, polyols, surfactants, amino acids, and antioxidants, to name a few. Finding the optimal combination components requires a methodical approach with matched analytics such as differential scanning calorimetry (DSC), dynamic light scattering (DLS), differential scanning fluorometry (DSF), and other product-specific techniques to down-select to a final formulation,” Chrimes states.

Lipid nanoparticles (LNPs) are emerging as a formidable technology that can facilitate molecule stability in biologic-based medicines. “LNPs facilitate the stabilization of the molecule due to their amphiphilic nature. This property allows large molecules to maintain their structure by interacting with either the polar head of the lipid or with the hydrophobic part of the lipid,” says Ailhas. Ailhas also points out that the variety of available lipids offers numerous options for LNP composition and stabilization strategies. Because of this variety, LNPs composed of different lipids are highly versatile, enabling them to stabilize various molecules, such as peptides or nucleic acids, with different charges.

Because LNPs can be used to encapsulate biotherapeutics, they protect the fragility of these therapies from accelerated degradation 

, Chrimes says. He explains that LNPs are biocompatible and naturally break down; however, the biodegradation needs to be timed to allow mRNA (as one example) to enter the target cells. “This type of delivery platform has accelerated the use of mRNA therapies. The control of lipid impurities generated during production of LNPs has also been reported to improve shelf life,” he adds.

LNP encapsulation shields molecules, such as mRNA, from temperature change, oxidation, and enzymatic degradation, Cao states. However, formulation development for mRNA-based therapies will still face the same challenges as mAb formulation development, Cao notes. “LNPs require different ratios of lipid components, such as ionizable lipids, PEG [polyethylene glycol]-lipids, cholesterol, and phospholipids, for their stability,” he emphasizes.

In addition to LNP technology, protein engineering and lyophilization are other techniques employed to facilitate the development of formulations that can offer better stability for temperature-sensitive molecules. “Development of stable products at ambient temperature is typically the preferred endpoint for biopharmaceutical formulations. However, it is not often obtained due to the physical forces experienced during transport, storage, and delivery,” says Chrimes. He explains that product liabilities can be removed in the primary sequence of a biotherapeutic, in some cases to eliminate the instability. He points to lyophilization as an often-successful technique to avoid colloidal instabilities that occur during shipment or long-term liquid storage.

Protein engineering and the use of machine learning, meanwhile, has been increasingly used to develop mAbs that are more thermostable, says Cao. Furthermore, the use of AI to analyze the local solvent-accessible surface area of the variable region of previously approved mAbs have been used to make predictions on aggregation, he notes.

“If proteins can get re-engineered, then the YTE mutations [M252Y/S254T/T256E] could be employed to achieve thermal stability,” adds Lim, who explains that once a molecule passes the discovery phase, a formulation scientist would then need to analyze basic characteristics of the molecule to identify its distinguished traits. “These analyses help us determine the onset temperature (Tonset) and melting point (Tm) of the molecules and provide us insight into how much the molecule can resist thermal stressors,” he says.

Furthermore, identifying pH/buffer levels is required to earn both thermal and chemical stability, Lim adds. “We can also add various saccharide excipients to formulations, such as sucrose and trehalose, to further enhance thermal stability by about 2–3 degrees Celsius. For sensitive/complex molecules [such as] antibody-drug conjugates, lyophilization may be the preferred technique to achieve maximum stability,” he states.

Meanwhile, other types of nanoparticles, such as polymeric ones, are being explored for thermal stability, says Ailhas. “Additionally, lyophilization combined with the appropriate selection of excipients should be investigated to enhance the stability of temperature-sensitive products,” she states. Unique excipients such as polyols, surfactants, amino acids, and antioxidants have been demonstrated to improve protein chemical stabilization and colloidal stability, Chrimes notes.

“Beyond LNPs, other type nanoparticle carriers such as polymeric nanoparticles, silica nanoparticles, and gold nanoparticles are being investigated for the capability to stabilize the molecules through encapsulation,” says Cao.

At a certain point, once a formulation strategy is in place, focus shifts to the drug delivery method. Drug delivery systems are usually considered in later clinical stages of drug development, notes Lim, who emphasizes that in early stages of drug development, getting a fast investigational new drug (IND) application approval is a high priority for drug developers. Speed in securing an IND approval grants a competitive edge to drug developers, especially in a drug discovery market that is dynamic. “The emphasis during the early stages is less on selecting a delivery system and more on obtaining reliable thermal and physical stability results for the targeted molecule,” he says.

Formulation development is usually initiated at the early stages of drug development as well, says Cao, who explains that early formulation development ensures the stability of the product and reduces the need for cold storage with, for example, techniques such as lyophilization. Formulations that result in stability can then inform the design of the delivery mechanism, he notes.

“A study for delivery can also be conducted first to test which delivery method (intravenous [IV] bag, subcutaneous injection, etc.) is effective for the therapeutical drug, followed by formulation development,” Cao also says. “Different modes of delivering the drug will require different drug concentrations. IV-bags do not require high concentrations of drug, but sub-cutaneous does. Challenges and limitations observed in the delivery mechanism can drive further optimization of the formulation,” he adds.

“In my opinion,” says Ailhas, “it is more beneficial to prioritize an optimized formulation that can function in a non-cold environment. This approach simplifies production and storage, eliminating the need to find specific drug product manufacturers capable of handling cold manufacturing processes, which can be challenging to locate.” Ailhas also points out that, although a formulation-first strategy may incur slightly higher costs at the beginning of a project, the long-term cost savings during the transition to good manufacturing practice manufacturing, either for clinical or commercial supply, will be significant.

Chrimes agrees that is it important to prioritize a formulation-first strategy: “Optimizing the formulation to limit the impact of temperature excursions would be the preferred development path for biotherapeutics,” he says, adding that every effort should be explored to limit potential liabilities as well as screen applicable excipients and explore presentations to “lock” the product in a stable state through techniques such as lyophilization, for example. Lyophilization removes water from sensitive products without damaging them. “Developing liquid formulations that are compatible with lyophilization can be an effective strategy for cost savings during clinical development where lyophilization cycles are deferred to commercial development,” he states.

Finally, what technical considerations remain to be solved that can conceivably facilitate a breakthrough to realistically realize temperature-stable biomolecules that no longer require stringent cold-chain handling?

Cao answers the question by pointing out that developing advanced stabilizers and excipients, as well as improving lyophilization and spray drying techniques, can aid in maintaining biological molecule stability without compromising efficacy or safety. Such advancements would allow biological molecules to be less dependent on stringent storage and transportation conditions.

“Don’t be afraid of lyophilization,” stresses Ailhas. “It can significantly help solve this problem, and it is not so difficult to manage, at least for mAbs. For mRNA vaccines formed with LNPs, the breakthrough will involve finding a way to remove water from the inner part of LNPs during lyophilization without damaging the LNP structure and ensuring they can return to their original form during reconstitution.”

Meanwhile, Chrimes says that the solutions are going to look different based on the modality, mechanism of action, mode of delivery, and more. Across the board, however, progress always starts with understanding what is causing the instabilities in a given biotherapeutic. “From there, we can work … to optimize not just for temperature stability, but shelf-life and other critical parameters that can impact commercial success. For the most part, it’s an ongoing process of optimization and refinement.” Chrimes adds that innovations in predictive modeling and machine learning could accelerate the development of unique excipients and formulation development.

“The key focus during the process development or clinical trial stages is to minimize the risk of failure,” adds Lim, who notes that, to minimize risk, a pre-molecular developability assessment is crucial. “Through this evaluation process, scientists can find any potential issues, such as lack of thermal stability, which can bring out exclusion or implementation of risk mitigation strategies for further development,” he says.

“This assessment can help researchers optimize strategies for thermal stability, including lyophilization as part of the formulation development process. Not only can this reduce the risk of failure but also save time during the development timeline,” Lim concludes.

®
Vol. 37, No. 9
October 2024
Pages: 12–14

When referring to this article, please cite it as Mirasol, F. Prioritizing Formulation Strategies for Temperature-Sensitive Biotherapeutics. 

Solving the challenge of better-stabilized temperature-sensitive biomolecules hinges on innovative formulation strategies.

Prioritizing Formulation Strategies for Temperature-Sensitive Biotherapeutics

Thermo Fisher Scientific announced on Oct. 7, 2024 that it is launching the Accelerator Drug Development, the company’s 360° contract development and manufacturing organization and contract research organization drug development solutions, at  at CPHI Milan 2024, happening in Milan, Italy from Oct. 8–10, 2024. The new services offering is part of the company's showcase at the conference, accordig to a company press release (1).

According to the company, Accelerator Drug Development will impact the pharmaceutical value chain for biotech and large pharmaceutical companies with its customizable manufacturing, clinical research, and clinical supply chain services for small molecule, biologics, and cell and gene therapies (1). The services are intended to assist customers at every stage of the drug development lifecycle.

“Thermo Fisher is transforming drug development and manufacturing processes to support customers as they move critical programs forward to tackle global health challenges with speed, quality and efficiency,” said Michael Shafer, executive vice president and president, biopharma services, Thermo Fisher Scientific, in the press release. “We connect the steps between discovery and delivery—from early testing and analytical support to clinical research design to trial conduct, clinical supply chain management, manufacturing and commercial scale-up—and we strive to ensure a seamless transition, mitigate risks, and accelerate innovation at every stage.”

The launch of Accelerator Drug Development comes on the heels of a busy summer 2024 for Thermo Fisher, which in June opened a new building at its good manufacturing practice (GMP) facility in Middleton, Wis., then announced the completion of its acquisition of Olink Holding, a Sweden-based provider of next-generation proteomics solutions, in July (2–3). The company has also expanded sites in Cincinnati, Ohio, and Bend, Ore., to enhance R&D, manufacturing, and testing capabilities for oral solid dose drug formulations (1).

Thermo Fisher can be found at Booth 7B18 at CPHI Milan. Sessions at which its experts are scheduled to be contributors include:

Thermo Fisher Scientific's launch of its Accelerator Drug Development follows expansions of sites in Cincinnati, Ohio, and Bend, Ore., earlier in 2024 to enable R&D, manufacturing, and testing.

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