Chronic illnesses or non-communicable diseases that affect the health of the global population are on the rise and, in turn, attributing to a large proportion of deaths and a higher economic burden year-on-year (1). To help combat these long-term illnesses, the bio/pharmaceutical industry has been investing in the development of novel therapies that are increasingly complex in nature.

“The complexity of drugs and their delivery has been increasing over the past few decades,” explains Peter Droc, vice president, head of Drug Product Services, Lonza. “Combined with a growing prevalence of chronic illnesses, this market evolution is putting more pressure on pharmaceutical manufacturers to develop novel therapies that might also have unique formulation needs.”

Many of these novel, complex therapies are biologic-based drugs that are delivered parenterally. “The injectable drug market is experiencing significant growth,” adds Gareth Jenkins, vice president, Science and Technology, Candidate Development, Quotient Sciences. “As a result, robust and efficient sterile manufacturing lines are essential across the manufacturing scales from the early phase to the commercial launch.”

“In its very nature, the aseptic processing market is bound to be less efficient than other market sectors,” remarks Nate Smart, process engineer, CRB Group. “[The] focus is delivering critical therapies to vulnerable patient populations in methods that bypass many of the body’s natural defense systems. Therefore, in contrast to other drug product markets (i.e., OSD [oral solid dosage]), the greatest care must be taken to ensure product quality is maintained.”

However, attaining high quality in aseptic processing is a time-consuming endeavor, as manufacturers need to pay close and accurate attention to adherence to strict guidelines, confirms Maximilian Kappler, process engineer, CRB Group. “The highest quality is imperative in aseptic processing to ensure product safety and effectiveness, making it non-negotiable to invest time for quality whether in design, processing, and product testing,” he states.

Regulatory authorities are scrupulous of all aseptic processing facilities and hold manufacturers to stringent standards, agrees Smart. “As a result, aseptic fill lines and their associated barrier systems are custom and costly,” he says. “In many facilities the aseptic filling equipment is the most expensive and longest lead equipment in the facility.”

As a result of these overheads, many manufacturers try to make their lines flexible so that the utility of their equipment can be maximized and new products, formulations, or formats can be added, Smart specifies. “The more products and formats you can run on a single line, the greater utilization and return on investment you can get out of your line,” he says.

“Despite these efforts, fill lines have notoriously low OEE [overall equipment efficiencies],” asserts Smart. “Many lines, even recent installations, have turnaround times of up to 24 hours or beyond. Any reusable critical contact parts (direct and indirect) must be washed and sterilized out of place and then installed in an aseptic manner.”

“The regulatory bodies are currently pushing for a continued reduced risk of contamination,” adds David King, senior director of Aseptic Processing at Samsung Biologics. “This push is focused on the removal of humans from the aseptic filling process, as humans are the primary source (directly and indirectly) of aseptic area microbial intrusions. The primary change is an industry-wide transition from open RABS [restricted access barrier system] filling systems to isolator contained filling systems.”

“The distinct advantage is that the environment around the open packaging container is sterilized (typically with vaporized hydrogen peroxide [VHP]) with the use of isolator systems,” King specifies. “Isolators cause a bottleneck in regards to small batches as the [process changeover times] can be complex with the requirement to reduce the risk of product contamination from hydrogen peroxide.”

“Ensuring sterility through the manufacturing process is critical and can lead to bottlenecks both for product release and during product changeovers or when there is a temporary issue on the line that is holding up production,” confirms Jenkins. A microbiology team can play a crucial role in helping to minimize such disruptions, he explains, as microbiology considerations can be incorporated at the process design stage and when optimizing operations.

“Sterility assurance is critical for quality control (QC) release of sterile drug products, and the industry-wide approach involves a two-week incubation period—both for testing the drug product and environmental monitoring of the facility,” Jenkins continues. “Numerous rapid sterility testing methods are now commercially available to help address this and can be applied in later stages of development.”

Additionally, supply chain disruptions, which have impacted many global industries over the past few years, can be challenging for aseptic processing, asserts Aimee Hodge, vice-president Business Operations, PCI Pharma Services. “Lengthy lead times for components and drug substance can be very challenging, especially for time critical projects,” she says.

“Delays or disruptions in the procurement of essential packaging materials can significantly impact production timelines and quality assurance efforts in aseptic processing,” states Kappler. “Addressing these supply chain challenges early in a project is crucial to maintaining the integrity and efficiency of the aseptic processing workflow.”

Improving the efficiency of the aseptic workflow is key to ensure critical therapies reach patients in a timely fashion while also being safe and effective. “Automating processes and bringing robotics into aseptic processing, together with single-use systems, [are] helping to streamline processes, minimizing human intervention and reducing the risk of errors or contamination,” emphasizes Hodge.

“Innovations in analytical testing such as rapid microbial and particle detection and rapid sterility testing can save valuable time for releasing drug product to clinical trials or commercial supply,” Hodge adds. “For example, with rapid sterility testing sample results are available in less than seven days, providing time efficiencies when compared to the 14-day incubation required for compendial sterility.”

However, regulatory requirements for viable particulate monitoring are based upon colony forming units, which “cannot easily be directly correlated with fluorescent particle counting results,” Smart asserts. “Manufacturers must be able to validate new methods against the existing standards to demonstrate the new method’s equivalence or superiority.”

Smart points out that rapid micro methods use biofluorescent particle counters in place of settle plates for viable particulate monitoring. “This technology has enormous potential to benefit manufacturers by providing real-time microbial data during aseptic processing,” he says. “Rather than waiting weeks to find out about EM [environmental monitoring] failures, operations can respond to issues as they occur and limit or avert resulting investigations and batch losses.”

“[Another] advancement that is now being more accepted by the regulatory bodies is the use of rapid microbial monitoring systems, [which not only] monitor particles within the filling environment, but also have the capability to determine if the particle is viable or non-viable,” King adds. “If the particle is viable, it is captured real time on a growth medium. This is reducing environmental monitoring excursion identification time frames.”

The presence of particles, subvisible or visible, in parenteral products causes delays and contributes to medicines shortages worldwide, specifies Belinda Holdsworth, strategic growth project lead, Drug Product Services, Lonza. “Having a robust, fast, and reliable method for visual inspection is therefore of utmost importance to drug developers, as well as to their patients,” she says. “An important trend in this space is the utilization of robotics and automated visual inspection (AVI) for defect and visual particle identification in drug products.”

“[Additionally,] the utilization of AI [artificial intelligence] for data analysis [of] image and pattern recognition significantly enhances the detection of difficult-to-inspect parenteral drug products and decreases false reject rates,” Holdsworth specifies. “Combined with an automated robotics approach, this robust process can eliminate the formation of air bubbles, thus greatly enhancing process timelines.”

Focusing on an engineering perspective, Kappler believes that industry will move toward more standardized processes. “In terms of smaller batches, the industry apparently tends to move toward established and validated components such as ready-to-use/ready-to-fill containers,” he says. “This [approach] allows [the manufacturer to] focus on the product core without investing too much time and effort on side processes.”

It has been a decade since ready-to-use pre-sterilized nests of vials were launched by glass pharmaceutical packaging specialist, Schott, explains Jenkins. “These [nests of vials] were designed in collaboration with filling-line manufacturers to eliminate certain processing steps, such as vial washing, drying, and sterilization at the drug product manufacturing line,” he says.

“The nested packaging also reduced incidents of glass vial damage due to vial-to-vial and vial-to-machine contact,” Jenkins states. “This [innovation] paved the way for advancements in robotics and automation, leading to a range of filling solutions, from small-scale support for early clinical supply to more robust systems capable of filling hundreds of vials per hour.”

Further innovation is being seen in the machine manufacturing space that reduces operator interactions, Kappler continues. “Advancements have been made surrounding environmental monitoring that is causing streamlining of operations of critical monitoring steps that ensure that the product remains aseptic throughout the process,” he confirms.

“Advancements in automation and robotics are revolutionizing aseptic fill/finish processing,” confirms Hodge. “From vial loading to stoppering, advanced robotics are streamlining and enhancing the efficiency of sterile fill/finish operations with reduced human interventions, minimizing contamination risks, while optimizing production speed and time to provide life-changing therapies to patients.”

Although, robotics still have some limitations, particularly in responding to faults, jams, and other unplanned events that are not programmable, Smart stresses. “In these cases, there is future potential for use of vision systems and machine learning/AI to provide robotics the capacity for responding to unplanned interventions without the risks inherent in human operations,” he says. “Manufacturers looking to maximize the benefit of robotics and other automation should seek to understand how to maximize flexibility in how they implement these solutions. A set of robotic tooling that is fit to a single format does not provide significant operational advantages over a set of single size format parts in a traditional line.”

“Time-to-market can mean the difference between having a viable product or the competing product taking the market,” emphasizes King. “The best way to reduce the time[-to-market] is to have all production activities at one site. If a company can produce the drug substance and drug product at the same site, product testing, storage, and release times can be greatly reduced.”

Managing to get a product to commercialization quicker also helps to provide patients with life-changing therapies in a better timeframe and ultimately improves their health outcomes, Hodge adds. “Biopharma companies can endeavor to accelerate the delivery of their drug product to patients by developing well defined processes and testing methods together with a product road map early in the drug development lifecycle to identify the steps to plan for to advance along the journey to commercial readiness and launch,” she says.

“Contract development and manufacturing organizations (CDMOs) also play a crucial role in supporting the accelerated supply of life changing therapies to patients by providing expert solutions for drug development, manufacturing, and clinical trial services,” Hodge specifies. “When partnering with a CDMO, to ensure a robust supply chain and support speed to market, it is vital that a preferred CDMO has global reach together with flexible, scalable solutions to support clinical trial from early phase through to commercialization.”

For Droc, the use of sterile, ready-to-use, and single-use packaging solutions is valuable when trying to increase product quality and patient safety, and also reduces time-to-market. “From the perspective of drug developers, these solutions can also accelerate time to market by streamlining operations and expediting product development,” he says. “For drug product manufacturing, pre-sterilized containers, such as vials, syringes, and cartridges, that are ready for filling are especially relevant.”

“Flexibility will be another key aspect of future time-to-market aspects,” reveals Kappler. “[Because] the variety of single products during clinic is high, systems that allow switching between different packaging materials, and quick change between different product set ups will increase usage and allow pharmaceutical manufacturers to quickly react on formulation in early phases.”

Although the benefits of reducing the time it takes to commercialize products are obvious, there is also a need to balance leveraging the use of innovative technologies with standard “tried-and-true” practices, Smart adds. “No one in pharma wants to jump in at the ‘peak of inflated expectations’ in the Gartner Hype Cycle, whereas waiting too long jeopardizes potential competitive advantages,” he says.

“While in many ways the [aforementioned] technologies are not new to the industry, their adoption still has far to go,” concludes Smart. “Courage is required to innovate, but the potential gains are significant. Given that regulatory agencies are themselves recognizing the benefits of these technologies, it seems that the way is paved for industry to reap the rewards of implementation.”

Hacker, K. The Burden of Chronic Disease. 

When referring to this article, please cite it as Thomas, F. Speeding Up Aseptic Processes. 

Articles

Cover Story

Technological advances are aiding manufacturers to streamline aseptic processes and improve time-to-market.

Speeding Up Aseptic Processes

Cell-culture media formulations comprise the ingredients required by cells to survive, grow, and function as needed for the intended cell-culture application. The exact formulation depends on the nature of the cell-culture process, the type of cell and specific cell line employed, and the class of biologic and target properties of the biomolecule being produced. Very small changes in the chemical composition of cell-culture media can cause significant changes in performance. Consequently, there are significant benefits to fine-tuning the media formulation for any given bioprocess.

Cells rely on many different chemicals and materials to stay healthy and active. Those used for biopharmaceutical manufacturing must also be able to express the target product in high yield and quality. To do so, cells require glucose and other carbon-based sugars as energy sources, amino acids, and peptides to support protein production; vitamins to act as cofactors for enzyme functions; lipids to serve as basic components for cell membrane construction; salts to achieve osmotic balance and enable many natural biologic processes; and trace elements used as enzyme catalysts, redox reaction components, and for other cellular functions, according to Kenneth Low, global head of media R&D in Lonza’s Bioscience.

In general, observes Kimberly Schrag, R&D manager, MilliporeSigma, the Life Science business of Merck KGaA, Darmstadt, Germany, modern cell-culture-media formulations are chemically defined (i.e., do not contain hydrolysates that often cause lot-to-lot variability) and do not contain any animal-derived components. She notes that in addition to amino acids, vitamins, trace elements/minerals, lipids, pH-buffering agents, and carbon sources, they can also contain recombinant growth factors, antioxidants, and some components for specific functions, such as poloxamer 188, which reduces shear stress for suspension cultures.

Amino acids, as well as serving as building blocks for proteins, are a major nitrogen energy source important for cell proliferation, adds Fei Chen, director of cell-culture process development at WuXi Biologics. Nucleotides and polyamines are also often included in media formulations to meet the needs of specific cells.

One of the keys to successful media formulation development, says Jana Mahadevan, R&D Manager, the Life Science business of Merck KGaA, Darmstadt, Germany, is maintaining the balance of these many components, as having the right composition directly impacts performance. “Imbalances in glucose and amino acid levels have been observed to result in heightened production rates of lactate and ammonium, affecting cell-specific metabolic processes,” she notes. Specifically, elevated glucose concentrations have been associated with the inhibition of cellular respiration and oxidative phosphorylation, a phenomenon known as the “Crabtree effect,” leading to increased glycolysis rates and sustained lactate production. “Meticulously balancing all media formulation ingredients, which provide vital nutrients to support cell growth, metabolism, energy production, and ensure osmotic balance and pH regulation, fosters consistent and reproducible cell-culture conditions,” Mahadevan concludes.

Generation of different types of biomolecules via cell culture involves different biological processes, which in turn dictate the needs of the cells involved. “Different bio-therapeutics, such as monoclonal antibodies (mAbs), other bioactive proteins, viral vectors [adeno-associated viral (AAV) and lentiviral (LV)], and exosomes have very different compositions and therefore may place very different nutritional requirements on the cells,” comments Josi Buerger, associate principal scientist involved in bioscience R&D at Lonza. For example, she notes that cells genetically engineered to produce mAbs and bioactive proteins may require higher amounts of amino acids in the culture media compared to cells bio-engineered to produce large amounts of exosomes.

There is a very important difference between processes producing a biopharmaceutical protein that gets produced by the cell (antibodies, enzymes, and various other proteins), viruses that get produced by the disruption of the cell (for vaccines or gene therapies), and if the cell itself is the product (cell therapy, but also non-pharma applications like cultured meat), Schrag agrees.

For instance, while media formulations for viral vector production are similar to those for recombinant protein and mAb production, the media for all products should be able to support rapid (low doubling time), high-density cell growth, and high-titer production, observes Mahadevan.

Consequently, media for AAV and LV vector production are formulated to be compatible with plasmid transfection for transient expression and thus have subtle differences from media for protein manufacturing, says Schrag. She goes on to note that even for AAV versus LV vectors, media must be formulated to meet different nutritional requirements because these two types of viral vectors differ significantly in composition and have different modes of infection (i.e., lytic vs non-lytic).

Not only do media formulations need to be adjusted to meet specific needs for growth and production, they must also be adjusted for the right product quality attributes (PQAs) to ensure both drug efficacy and safety, according to Zhou Hang, senior vice-president and head of bioprocess research and development at WuXi Biologics.

For most therapeutic proteins (mAbs, bispecific or multi-specific antibodies, antibody fragments, antibody-drug conjugates [ADCs], etc.), purity is always important due to its potential impact on drug safety, efficacy, and stability. Meanwhile, post-translational modification attributes, such as glycosylation profiles, and charge-variant distributions may be related to drug potency. “Each molecule may have different challenges in PQA tuning. For example, maintaining a higher purity for a bispecific or fusion protein is usually challenging, and thus more attention needs to be paid to media and process development. For biosimilars, PQA profiles must be finely tuned to demonstrate comparability to their originator molecules,” Hang explains.

Another reason different media formulations are required relates to the use of different types of cells. Different cell types may have very different basal nutritional needs that must be considered within the context of maximizing cell growth, product yield, stability, quality, and correct protein folding/conformation, remarks Alexis Bossie, director of media R&D in the Bioscience business at Lonza.

There are different nutritional needs for primary cells vs. immortalized cell lines, according to Bossie. “Primary cells from human or other animal sources require serum, growth factors, and any other support that in vivo systems deliver. In vitro culture of primary cells will require all these elements to be present in the culture medium,” she says. Immortalized cell lines, however, can propagate in the absence of high concentrations of serum, cytokines, and other growth factors in the culture media.

For biopharmaceutical proteins, Chinese hamster ovary (CHO) cells are well established, and media and processing technologies are mature, according to Schrag. Consequently, high-performing media is readily available off-the-shelf.

The variety of production hosts for vaccines and gene therapies has expanded and now includes HEK293 and other mammalian cells, as well as insect cell lines. “For these platforms, media and feeds exist but the technology is not as mature as for CHO cells,” says Schrag. For example, she notes that some of the commercially available media are not chemically defined or even require animal-derived components.

With cell therapies, it is important to maintain the quality parameters of the cells to ensure they retain their therapeutic potential, as the cell is the product, Schrag observes. Unlike for other biologics, several growth factors are needed, creating an interesting challenge for media development. “Only when these molecular biochemistry needs are met, the classical media development and optimization of nutrients can be initiated, and constant checks on the cells’ quality are needed,” she explains.

Variability within cell lines derived from the same lineage is an added challenge, according to Mahadevan. “The choice of specific cell line used for producing different biomolecules can impact the media requirements, as each cell line may have specific nutritional needs and sensitivities,” she explains. As is the case for different proteins, media formulations must often therefore be tailored for specific cell lines to ensure proper post-translational modification and folding to support production of stable products with therapeutic efficacy.

Cell-culture processes can also be run under several different conditions, all of which impact the optimal composition of media formulations. Adherent vs. suspension and fed-batch vs. perfusion are just two examples.

Adherent media formulations tend to be leaner because the cell densities are lower compared to suspension cultures, and adherent media contains higher levels of components that enable attachment, like Ca/Mg, says Schrag. In addition, while there are some chemically defined, animal-component-free media for attached cultures, she points out that many attached processes (e.g., virus production for vaccines) still use a basal medium plus fetal bovine serum (FBS).

Suspension media are more concentrated than adherent media because suspension processes operate at higher cell densities. They also often contain components such as poloxamer 188 and anti-clumping agents, according to Mahadevan. Perfusion media are even more concentrated than conventional suspension media and contain much higher levels of components that are exhausted during growth and recombinant protein production, including amino acids and glucose.

In addition to these reasons for modifying media formulations, there are other drivers for adjustment to meet specific process requirements. Even for the same cell line, says Chen, the transfection medium, cell-passage medium, and production (fed-batch or perfusion) medium are quite different. Transfection media must be formulated considering how they can impact the transfection process. “Metal ions such as iron (III) are reported to inhibit transient transfection due to their effect on polyethyleneimine transfection reagents,” she observes. For cell-passage processes, cost-effective formulations that support desirable cell growth rates and ensure clone stability take precedence.

Nutrient-rich basal media, meanwhile, are typically preferred for both fed-batch and perfusion production processes, according to Chen, although for fed-batch processes, supplemental feeds are used to boost cell growth and enhance productivity, while for perfusion processes more attention is paid to maintaining cell viability. “Fed-batch platforms often require feeds to keep cell growth, titer, and product quality high, whereas perfusion/continuous processing platforms require media formulations that will keep the cells at a steady state for longer periods of time once the desired growth to a critical mass is achieved while maintaining high product quality,” Bossie agrees.

In addition to the complexity created by diverse molecule formats, cell types, and process conditions, cost considerations are another big challenge to media development. “The goal is to develop the best media formulation that maintains high cell growth, product titer, stability, and quality (correct protein folding, assembly, and glycosylation) at the lowest possible cost,” states Low.

For instance, he notes there is a definite trend toward developing fully chemically-defined media formulations with components that are of non-animal origin and are devoid of expensive ingredients such as growth factors or hormones. He adds, however, that it can be difficult to get cells that exhibit robust growth and produce bioproducts at high titers while maintaining high product quality and stability to be less dependent on these expensive components.

Development of a small-scale and high-throughput model for media development is also crucial, Hang observes. “Such models need to be sufficiently representative while also cost-effective,” he emphasizes. As an example of a problematic approach for intensified perfusion processes, Hang points to the spin tubes widely used for media screening and optimization. “These tubes show some limitations in process control and mass transfer in certain culture modes, including perfusion, and therefore are not adequate small-scale models for media formulation development,” he says.

Successful media formulation requires a deep understanding of the specific biologic drug substance, cell type/line, and process involved, including metabolic demands and support for specific-post-translational modifications. “This knowledge is employed using statistical experimental design methods to systematically evaluate and optimize media components and conditions,” says Schrag.

Computational and experimental tools used to attain the desired cell growth, protein expression, and productivity include design-of-experiment (DoE) studies with systematic varying of media component concentrations, computational modeling (multi-factorial analysis) for optimizing nutrient utilization, machine-learning algorithms for predicting complex interactions between media components and cellular responses, high-throughput screening for identifying optimal conditions, development of media supplements and feed strategies, and other mathematical models for predicting optimal compositions, according to Mahadevan. “Employing a combination of these tools and approaches enables systematic optimization tailored to specific cell lines, processes, and desired outcomes,” she concludes.

For instance, starting from media screening, Chen explains that key components can be identified and optimized using various statistical analysis methods. In this manner, selected key subgroups or components in the formulation can be identified and their concentrations optimized based on the statistical data. “Due to the complexity of cell-culture-media-formulations, however,” Fei cautions, “it is not practical to use statistic approaches to simultaneously optimize all components, as the time and workload involved would be too onerous.”

The best strategy for media formulation development for biologics manufacturing often involves a systematic and iterative approach tailored to the cell line’s specific needs, according to Buerger. Media development should also focus on ease-of-use, adds Ioanna Zormpa, associate principal scientist supporting biologics media R&D in Lonza’s Bioscience business.

There are four main steps in media formulation development, including basal media formulation, media optimization, performance evaluation, and scale up. The starting basal media formulation should include sufficient components for the specific application, as well as supplemental feeds if appropriate, with regulatory requirements for sourcing media components taken into consideration, Buerger says.

Optimization studies should be in concert with process development to facilitate ease of integration into downstream manufacturing processes. Evaluation of the selected optimized media formulation for growth rate, productivity, and product quality requires appropriate media analytical capabilities, some of which may require development of new analytical methods. Buerger therefore recommends developing an overall strategy adaptable to these differing needs. Scale-up studies ensure that performance of the cell line and media formulation remain consistent as processes move from the lab to the production plant.

The development of media formulations that are stable over time is also essential for maintaining consistent cell-culture performance and product quality. Therefore, conducting appropriate expiry studies to determine media shelf life is another key part of this process, according to Zormpa, as it helps to ensure effectiveness and safety over a specified period.

Throughout these steps, the production of easy-to-use media products should be kept in mind, as they can simplify the manufacturing process, reduce the potential for errors, and increase efficiency for end users, Zormpa adds. As an example, she points to single powder formulations, which simplify media preparation, lower the risk of component omission, and enable straightforward water reconstitution protocols, make them a practical and efficient choice for biologics manufacturing.

“These practices collectively contribute to a more streamlined, reliable, and effective media development process,” Zormpa concludes.

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

When referring to this article, please cite it as Challener, C.A. Addressing the Complexities of Media Formulation Development. 

Upstream Processing

Using a multi-pronged strategy to find optimum, tailored formulations is best.

Addressing the Complexities of Media Formulation Development

Continuous processes continue to advance in bioprocessing. In downstream processing, continuous chromatography is gaining traction in the purification of protein therapeutics. As the biopharmaceutical industry strives to demonstrate, continuous manufacturing offers the benefits of higher productivity and better product quality than traditional batch manufacturing. In addition, continuous processing intensifies production and lowers capital costs, while enabling better control (1).

Although significant strides have been made to advance continuous upstream processing, downstream processing has not advanced at the same pace. However, according to Gerstweiler et al., “Research on continuous chromatographic purification for antibodies is maturing, and work has commenced on other unit operations and on process system integration” (1).

Somasundaram et al. had earlier noted that “Rapid advances in intensifying upstream processes for biologics production have left downstream processing as a bottleneck in the manufacturing scheme” (2). But to increase efficiency and promote flexibility, biomanufacturers are looking to implement continuous downstream process development. However, despite successful laboratory trials, it has remained difficult to achieve continuous downstream processing at commercial manufacturing scale.

Perhaps another challenging aspect with continuous downstream processing is the ability to implement process control strategies. Advances in process analytical technologies and modeling techniques, however, may provide solutions to improving chromatography control for industrial-scale manufacturing (3). These advances offer opportunities to enhance process robustness, increase productivity and move towards real-time release testing.

It has been previously noted that process optimization and controlling the steps in downstream chromatography contributes to increased consistency of product quality as well as better management of process variability. In addition, control of the downstream processing steps can lead to cost reductions (3). However, the biopharma industry needs to enhance monitoring and control strategies to realize these benefits as current chromatography control strategies remain limited, leading to sub-optimal processing operations. Ultimately, this limitation in control strategies leads to delays in purification for new therapeutic molecules (3).

®, Christine Rozanas, PhD, global product marketing manager at Cytiva, discussed the advantages and persistent challenges with implementing continuous downstream chromatography.

 What is continuous chromatography, and how has it improved downstream bioprocessing?

 Continuous chromatography differs from traditional ‘batch’ chromatography in two key ways: the chromatography system can be loaded continuously, which is a useful approach when the purification step is connected to an upstream perfusion or fed-batch bioreactor, and, by having two or more columns in the loading zone, you can overload the first column [because] any product/target molecule that is breaking through is being captured by the second column in the loading zone. When used with a perfusion or fed-batch bioreactor, continuous chromatography can help reduce the amount of resins and buffers needed. It also provides greater flexibility for processes and uses a smaller footprint in a manufacturing facility.

 What are the benefits of using a continuous chromatography process in downstream bioprocessing?

 Compared with batch operations, continuous chromatography can offer significant productivity gains and increased production capacity, for example, through increased utilization of chromatography resin binding capacity, reduced buffer usage, and smaller column size.

 What have been the major drawbacks or challenges to using continuous chromatography in downstream bioprocessing?

 Despite the benefits, the industry has been to slow to adopt continuous chromatography in downstream bioprocessing for a number of reasons. It can be a more complicated set up compared to a traditional batch process, and, while batch chromatography is done over a single shift or sometimes over 24 hours, the larger volumes processed continuously from perfusion or fed-batch bioreactors must be processed over several weeks or up to a month. [This] can make it more difficult to trace any potential issues that might occur. While regulatory agencies express interest in supporting the use of continuous processing, many companies continue to develop batch purifications first, before evaluating whether a continuous chromatography process would significantly reduce production costs or increase productivity and output.

 How does multi-column continuous chromatography work, and in what scenarios would a biomanufacturer opt to implement this?

 For multi-column chromatography, the large capture column is split into multiple smaller columns that alternate in binding and elution operations in multiple cycles to process the feed from the upstream bioreactor. It can be a great option when working with sensitive molecules, where continuously removing the molecule from a potentially harmful environment into the first purification step can be the only viable option. In scenarios where there is a limited product demand (in kg), continuous chromatography can also allow significant reduction of the needed resin volume through running a higher number of cycles in the available time and thus reducing the column size.

 What key innovations have been instrumental in promoting the use or adoption of continuous chromatography?

 [Some] continuous chromatography systems use controls (e.g., dynamic ultraviolet control, as with Cytiva’s systems) that enable real-time monitoring and control of the process. In Cytiva’s case, its system can adjust accordingly to changes in feed concentration, for example, to optimize processing of the perfusion output. Also, pre-packed columns have provided greater consistency and quality of the internal conditions of each column so that the feed is processed in the exact same way in the alternating columns during the continuous run.

1. Gerstweiler,L.; Bi, J.; Middelberg, A. P. J. Continuous Downstream Bioprocessing for Intensified Manufacture of Biopharmaceuticals and Antibodies. 

, 116272. DOI: 10.1016/j.ces.2020.116272
2. Somasundaram, B.; Pleitt, K.; Shave, E.; Baker, K.; Lua, L. H. L. Progression of Continuous Downstream Processing of Monoclonal Antibodies: Current Trends and Challenges. 

 (12), 2893–2907. DOI: 10.1002/bit.26812
3. Armstrong, A.; Horry, K.; Cui, T.; et al. Advanced Control Strategies for Bioprocess Chromatography: Challenges and Opportunities for Intensified Processes and Next Generation Products. 

, 461914. DOI: 10.1016/j.chroma.2021.461914

Feliza Mirasol is the science editor for 

®
Vol. 37, No. 6
June 2024
Pages: 21–22

When referring to this article, please cite it as Mirasol, F. Evaluating the Use of Continuous Chromatography. 

Downstream Processing

The biopharma industry is looking at continuous chromatography for sensitive molecules.

Evaluating the Use of Continuous Chromatography

Prefilled products, particularly prefilled biologics, are a growing sector in the pharmaceutical industry. “Prefilled syringes and pens offer precise dosing, reducing the risk of errors during administration. They also eliminate the need for reconstitution, making them more convenient for patients and healthcare professionals,” explains Ward Smith, director of marketing and business development at Keystone Folding Box Company, a manufacturer of paperboard packaging. Prefilling also improves product/patient safety because less handling means less chance of contamination.

Prefilled biologics address the growing demand for personalized medicine, homecare, and self-administration options, as well as the desire for an improved patient experience. “This allows patients to continue their treatment in the comfort of their homes while reducing the costs for the healthcare system as fewer trips to a hospital or doctor are needed,” notes Tom Van Ginneken, product manager of polymer solutions at SCHOTT Pharma, a manufacturer of containers used for prefilled biologics and other pharmaceutical products.

The latest innovations for prefilled biologics, including pens, auto-injectors, and on-body delivery systems, accommodate higher volumes, more viscous fluids, a proliferation of longer-acting injectables with longer dosage intervals (weekly, monthly, or even longer), smaller batches, and the ever-present need to reduce healthcare costs. As a result, manufacturers are focusing on user-friendly device designs with features such as ergonomic grips, intuitive mechanisms for injection, and safety features to prevent accidental needlesticks. One example, the Aidaptus auto-injector, developed by Owen Mumford and Stevanato Group, can accommodate a variety of drug viscosities. For added flexibility, the two-step, single-use auto-injector is compatible with 1-mL or 2.25-mL prefilled glass syringes (1).

“Some prefilled biologics packaging now integrates digital technologies, such as Bluetooth-enabled devices or smartphone apps,” reports Smith. These connected devices can track dosing history, provide reminders, and transmit data to healthcare providers, improving adherence and patient monitoring.

Finally, sustainability is an increasingly important consideration. Therefore, manufacturers are exploring ways to minimize environmental impact by moving away from non-renewable materials, optimizing packaging size and weight, incorporating recyclable, recycled-content, or biodegradable materials, and maximizing reusability.

Primary packaging developments include ready-to-use (RTU) containers and designs to address higher viscosity products, larger syringe volumes to meet the dosing requirements of longer-acting biologics, cryogenic storage and distribution, greater focus on self-administration and patient comfort, and container enhancements that minimize the risk of product/package interactions. “This minimized interaction risk ensures that the biologics maintain their efficacy and safety profiles,” says Van Ginneken.

With sensitive substances and challenging environmental requirements, biologics place special demands on primary and secondary packaging. Many APIs are light-sensitive, and the large, complex molecules in many new molecular entities are sensitive to oxygen. Cell and gene therapies as well as messenger RNA (mRNA) substances must be stored and transported at extremely low temperatures. Thus, syringe development has focused on designs capable of withstanding low-temperature conditions without losing functionality or container closure integrity. One example, the TOPPAC freeze prefilled syringe from SCHOTT Pharma, is designed for drugs that require storage and transport at temperatures approaching -100 °C. Another option from SCHOTT Pharma, EVERIC freeze vials, are available in sizes from 2–30 mL and can be supplied RTU. Strength-optimized geometry, designed into the structure of the vials, reduces the risk of breakage (2).

As drug products have moved toward more complex proteins and longer intervals between injections, “primary packaging needs to evolve,” says Daniel Martinez, product manager at Stevanato Group, a global provider of drug containment and delivery solutions. This evolution requires higher levels of inertness to minimize chemical interactions, which can cause aggregation that could trigger an immune response in a patient.

Glass continues to dominate, but cyclic olefin copolymer (COC)/cyclic olefin polymer (COP) syringes are well-established alternatives due to their break resistance, lighter weight, lower chances of protein absorption, and compatibility with cryogenic conditions. Unfortunately, COC/COP’s barrier properties may not be sufficient for oxygen-sensitive biologics and

biosimilars. However, “… specialty labels with built-in barrier functions can overcome this challenge,” reports Corinna Endres, product manager drug protection solutions at Schreiner MediPharm, a Germany-based provider of functional labels. The company’s Gas-Protect-Labels can reportedly reduce oxygen transmission without changing processes or primary packaging. One Gas-Protect-Label design covers the syringe barrel. Another design, the Syringe-Closure-Wrap, enhances barrier protection by wrapping like a second skin around the syringe barrel and cap and sealing at the upper end. An integrated first-opening indication on the Syringe-Closure-Wrap also provides tamper evidence. Label structures can be customized for the barrier needs of the API in the syringe.

Schreiner Medipharm also offers a label for prefilled cell and gene therapies and mRNA substances stored and transported under cryogenic conditions. The Freeze-Light-Protect label reliably adheres to COC syringes at sub-zero temperatures and provides customizable light protection ranging from blocking pure ultraviolet (UV) rays, to blocking UV rays and blue light, to providing a total light barrier. Freeze-Light-Protect labels can be equipped with integrated functionalities such as detachable documentation labels, an inspection window for checking the syringe content in true color, and a graduation for exact dosing when performing the injection. “With the help of this multi-functional label solution, product and patient safety can be enhanced,” says Endres.

Although 1 mL remains the most common volume for auto-injectors, the range of prefilled syringe and cartridge sizes is expanding to include 2.25-, 5-, 10-mL, and greater volumes needed to handle more viscous products, higher volume injections, and products that can’t be concentrated and are destined for on-body delivery systems. SCHOTT Pharma, for example, has added a 5.5 mL size to its syriQ BioPure glass syringe family. Applications include larger volume medications for treating cancer and autoimmune diseases.

It’s also been necessary to optimize mechanical performance to accommodate higher impact forces without breakage. “More viscous products require higher injection forces,” explains Martinez.

The development of device platforms, like the two-step YpsoMate injector from Ypsomed or the Molly auto-injector from SHL Medical, is streamlining the introduction of combination products. Instead of performing Phase I and II clinical trials in vials and moving to a prefilled syringe for Phase III and commercial product, developers are adopting prefilled syringes as early as Phase I or II. “This … allows for earlier data gathering for the prefilled syringe presentation, and perhaps even more importantly, one can obtain valuable stability information at a much earlier stage,” says Justin Schroeder, global vice president, technical sales at PCI Pharma Services, a contract development and manufacturing organization (CDMO). Being able to include real-world data in regulatory filings can expedite regulatory approvals and shorten time-to-market.

The earlier transition to a prefilled syringe also helps ensure the patient self-doses properly, prevents medication errors, mitigates potential risks, improves patient adherence and persistence, and increases patient education and health literacy. “Well-orchestrated and integrated device, packaging, and labeling strategies can succeed on all [these fronts] if planned and executed synchronously,” explains Schroeder.

A customizable on-body delivery system, the Vertiva platform developed by Stevanato and Thermo Fisher Scientific, consists of a single-use pod with prefilled and pre-loaded 3-mL cartridge and a multi-use, smart controller. Stevanato supplies pre-sterilized EZ-fill cartridges and assembly equipment, while Thermo Fisher provides fill/finish and final assembly services (3). The highly flexible programmable device can deliver micro-precision basal doses or full-content bolus injections and is suitable for a wide range of therapies. It also accommodates volumes greater than 3 mL and offers Bluetooth connectivity for digital health applications. Reusability helps address sustainability concerns (4).

The primary trends on fill/finish lines for biologic products are the growing adoption of RTU containers, higher levels of automation and flexibility, and increased reliance on CDMOs.

RTU containers result in a more flexible, leaner operation and improve contamination control. For example, EZ-fill Smart containers from Stevanato now rely on thermally welded polymer film tray lidstock. The change from traditionally used Tyvek lidstock requires minimal modification of the fill/finish line and eliminates any fiber or glue residue contamination associated with the removal of the Tyvek material.

Flexibility, especially to handle the smaller batches needed for clinical trials or personalized medicine, is becoming an essential attribute of the fill/finish process. One filling system installed at PCI Pharma Services, the SA-25 aseptic filling workcell from Cytiva, handles vials or syringes. “These advanced robotic isolator systems are Annex 1-compliant and offer very small line losses, [and] are attractive for those developing high-value drug substances such as biologics and antisense oligonucleotides,” reports Schroeder.

When automating, some companies transition to fully automated systems, while others take a semi-automated approach. One semi-automated example, a custom-built benchtop draw filler from Turbofil Packaging Machines, offers a system that is more accurate and less labor-intensive than manual filling. Plus, it fits under a laminar flow hood, making it suitable for aseptic filling. The machine fills the syringes through the tip directly from intravenous bags via a disposable fluid path. This “saves time and resources while reinforcing sterility,” explains Deborah Smook, vice-president of marketing and business development at TurboFil Packaging Machines. Draw-filling not only enhances accuracy but also improves operator safety, simplifies changeover, accommodates a variety of syringe sizes and product types, and eliminates the need to clean a metering device.

For fully automated operations, a servo-driven, intermittent-motion TipFil monoblock model loads, fills, caps, inspects, and labels syringes and prints lot number and expiration date and other information such as QR codes or usage instructions at up to 50 pieces/min. Interchangeable pucks accommodate syringe sizes from 1–60 mL, and the machine can handle a wide range of product viscosities (5).

However, Smook warns, “… automation is rarely as simple as devising a machine to replace a manual process. Adjacent issues such as sterility and inspection, which can become more challenging at higher line speeds, also must be addressed.”

Automated inspection needs can be met by the high-speed Visual Rotating Inspection (VRI) series from Antares Vision Group. Capable of checking liquid-filled glass or plastic containers at speeds up to 400 containers/min, the machines combine sophisticated particle and cosmetic detection with closure integrity verification. VRI units can inspect liquids with viscosities from water-like to gels and emulsions, as well as lyophilized and powder products. Vials also are inspected for cosmetic defects related to crimping quality, flip-off color, neck-shoulder sidewall flaws, and stopper position (6).

Overall, drugs are becoming more complex and thereby more sensitive. Thus, “… the prefilled biologics packaging industry is characterized by ongoing innovation aimed at enhancing safety, convenience, and patient experience, while also addressing regulatory compliance and sustainability concerns,” says Smith.

“It is therefore essential for pharma companies and containment suppliers to work hand-in-hand from the beginning to ensure that the medication is stored and administered safely,” concludes Van Ginneken.

1. Stevanato Group. Aidaptus Manufacturing. 

 (accessed March 4, 2024).
2. SCHOTT Pharma. Protecting Drugs at Deep-Cold Temperatures: SCHOTT Pharma Launches New Vials for mRNA and Gene Therapy. Press Release. Jan. 24, 2024.
3. Stevanato Group. Stevanato Group Collaborates with Thermo Fisher Scientific to Bring Its Innovative On-Body Delivery System Platform to Market. Press Release. March 21, 2023.
4. Stevanato Group. Vertiva. www.stevanatogroup.com (accessed March 4, 2024).
5. TurboFil Packaging Machines. TurboFil Introduces Fully Automatic Version of Popular Syringe Filling System. Press Release. Dec. 5, 2023.
6. Antares Vision Group. At INTERPHEX, Antares Vision Group to Showcase Automatic Inspection Machines for Vials, Pre-filled Syringes, and Other Small-Volume Containers. Press Release. Feb. 13, 2024.

When referring to this article, please cite it as Forcinio, H. Innovations in Prefilled Biologics. 

Manufacturing

Packaging and packaging line innovations are addressing the industry’s growing use of biologics.

Innovations in Prefilled Biologics

The COVID-19 pandemic has given the bio/pharma industry lessons on being prepared to manufacture parenteral formulations for human use in emergency situations. Vaccines, remdesivir injection and amphotericin injection, are such recent examples. The challenge then becomes having enough flexibility in the manufacturing lines to facilitate manufacture of large volumes of unit doses of injectables as may be required with shorter changeover times, while assuring compliance and patient safety from the product contact parts used in the manufacturing line. Single-use manufacturing solutions provide the flexibility in terms of batch sizes, fill volumes, and ease of manufacturing, especially in terms of manufacturing different products in the same line with minimum line changeover requirements. This article reviews and elaborates the initial risk assessment process for extractable and leachables arising from single-use product contact manufacturing parts as per 

) chapters <665> and <1665>, which go into effect May 1, 2026 (1,2). The determined risk can then be used to build the level of extractable profiling and process equipment-related leachables (PERLs) followed by assessing the patient safety for regulatory submissions.

Manufacturing systems used for biopharmaceutical drug substances or drug products can be partially or completely constructed from plastic materials. During the manufacturing process, the drug product and its components could directly come in contact with the plastics and potentially interact with one or more of the manufacturing system’s product contact components. The substances that leach from a product contact manufacturing component could become incorporated into the process stream. If these leached substances persist in the process stream and continue through the process operations, they could accumulate in the drug substance or drug product as PERLs (1,2). When present in the drug product, these PERLs can potentially alter the critical quality attributes of the drug product, such as its stability, efficacy, and safety. 

 <665>, Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products, addresses these interactions by providing a risk-based means for chemically characterizing and qualifying plastic components used to manufacture biopharmaceutical drug substances and pharmaceutical and biopharmaceutical drug products. 

 <1665>, Characterization and Qualification of Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products, communicates the key concepts behind and provides additional information and guidance regarding the applicability and the application of 

 <665>. The authors discuss a practical approach by which 

 <1665> can be applied in a step-by-step manner for the risk assessment, using a case study involving the manufacturing process for remdesivir injection and single-use technologies (Merck).

The testing of a product contact component is driven by the risk that the component may be unsuited for its intended use based on chemical or biological effects attributed to PERLs (1). As such risks increase, the degree of required testing also increases. The risk assessment process is accomplished by a risk evaluation matrix that establishes the appropriate contributors to the dimension of risk, provides a quantifying means for the risk in each dimension, quantifies the total risk for each dimension, and links the quantified risk to an appropriate characterization strategy.

The risk evaluation matrix evaluates the dimensions that address the risk that a product contact part will be leached by a process stream to the extent that the extractables may be impactful on the final product. As shown in 

, the dimensions include the duration of product contact, temperature during product contact, chemical composition of the product stream, and the nature of the material with which the product contact component is constructed. The risk evaluation matrix will then evaluate each dimension separately and assign a level of risk based on certain measures relevant to each dimension. 

The risk evaluation of condition and duration of contact with the process stream addresses the driving force for the leaching. Depending on the duration of contact, the dimension has been divided into short term (<24 hours, Level 1), intermediate (1–7 days, Level 2) and long term (> 7 days, Level 3).

The risk evaluation of temperature of the process stream or the component during the contact duration drives the magnitude of leaching. Depending on the temperature during contact time, the dimension has been divided into low (refrigerated, Level 1), intermediate (ambient, Level 2) and high (elevated, Level 3).

The risk evaluation of the chemical nature of the contacting process stream addresses the process stream’s leaching power. The chemical composition of the process stream in terms of “aqueous”, “somewhat organic”, and “highly organic” are further discussed in 

In case the process stream contains multiple solubilizers, such as a protein solution with a surfactant, the process stream is risk-assessed as a compounded risk associated with the individual agents.

The risk evaluation of the chemical and physical natures of the contacted component addresses the component’s propensity to be leached. The risk dimension for “the chemical composition of the component” can be identified as low, intermediate, or high based on the composition or physical properties of the component (2). By composition, the risk dimension can be identified based on the % level of additives in the plastic material. The component is considered to be low-risk if the plastic additives are ≤0.1% by weight, intermediate risk if the plastic additives are between >0.1% and ≤1% by weight, and high risk if the plastic additives are >1% by weight. By physical properties, the risk dimension is considered to be high risk if the component is sterilized by irradiation or at high temperature.

The component that is being assessed for risk is rated for risk dimensions as per 

. The rating levels are assigned as 1, 2, or 3 in each of the risk dimensions. A numerical risk sequence is obtained as a result of the four assignments. If the level of risk is high for all four dimensions, then the numerical risk sequence will be 3333. The numerical order of the risk sequence values is largely inconsequential, and proper use of the numerical risk sequence requires that the sequence be given in order of decreasing digit values. Considering all the possible outcomes of the above application of the risk evaluation matrix, links between the numerical risk sequence and the characterization level have been proposed by 

Once the characterization level is determined, the risk evaluation process should then consider the mitigation factors that can be used to adjust the characterization levels. After the component has been evaluated, the downstream process operations (such as the clearance step—for its ability to either eliminate, remove, or clear PERLs from the process stream or dilute them to an extent that the safety of the manufactured drug product is not affected since the extractables will not be present in the drug product as leachables post-this step) can be used to mitigate the risk by 1 level, and the level of characterization required. If the component is subject to flush or rinse as a part of the manufacturing process and the flush/rinse waste does not go into the main process stream, then this flushing or rinsing has an ability to reduce the number and amount of extractable in the final drug product. If the reduction or elimination of extractables has been established with the flushing or rinsing step, then the material reactivity risk level can be reduced by one level, for example a “high risk” component can become an “intermediate risk” component. Process dilution to a certain extent can be considered in terms of its ability to reduce or mitigate the effect of the extractables and can be used to mitigate the risk by 1 level. Clinical use can be considered as a mitigation factor for non-parenteral dosage forms clubbed with their daily dose volume and the duration of the clinical use. If the sum of mitigation factors that apply is 0, then there is no adjustment in the characterization level. If the sum of mitigation factors that apply is 1, then the characterization level established by other dimensions is reduced by one level of testing (example level B testing is reduced to level A testing). If the sum of mitigation factors that apply is 2, then characterization level A is applicable in all circumstances.

The next example looks at the risk evaluation process in a step-by-step manner for the risk assessment of the sterilizing grade filter used as a part of a single-use assembly (Mobius, Merck) in the manufacturing process of remdesivir injection.

 shows a representative schematic of the single-use assembly used in the fill/finish of a biopharmaceutical drug product in this case study (3). In this example, the assembly has a polyvinylidene fluoride filter (Durapore, Merck) along with silicon tubing and connectors as the product contact parts. The assembly is sterilized by gamma irradiation. 

Remdesivir for injection, 100 mg/vial, is a preservative-free, white to off-white or yellow lyophilized solid. The concentrate for solution for infusion contains 100 mg remdesivir that is to be reconstituted with 19 mL of sterile water for injection and diluted into intravenous (IV) infusion fluids prior to IV administration. Remdesivir injection is produced by aseptic filtration. The process steps include dissolution of betadex sulfobutyl ether sodium in water for injection, pH adjustment and dissolution of remdesivir, dilution with sufficient quantity of water for injection, pH adjustment (pH 3.0 to 4.0), bioburden reduction via filtration, sterile filtration, vial filling, stopper insertion, lyophilization, and capping. The product before lyophilization contains remdesivir 5 mg/mL, betadex sulfobutyl ether sodium 300 mg/mL, hydrochloric acid and/or sodium hydroxide in a quantity sufficient enough for pH adjustment, and water for injection. The liquid after sterilization by sterile filtration is filled into 20-mL vials under aseptic conditions before being subjected to lyophilization. Water for injection used as a base is removed during lyophilization (4,5). For this case study, it is assumed that the manufacturing batch size is 500 L and the maximum contact time of the final sterilizing grade filter is 12 hours at a temperature of 22.5±2.5 °C. It is also assumed that there is no flushing or rinsing during the manufacturing process.

Applying the risk evaluation matrix from 

 to this case study results in the risk level rating shown in 

The risk sequence will therefore be 3211. As per the manufacturing process provided, there are no steps that can be considered for risk mitigation, and hence the risk mitigation factor is 0. The risk level for level of characterization will be Level A or B. Applying the clauses as defined in 

, it can be determined that a characterization of Level B will be required for this component.

Following the risk assessment, the next step will be to design and execute the extractable test corresponding to Level B, as per 

 chapters <665> and <1665>, generating the extractables profile and PERLs. Extractables will be further evaluated for patient safety assessment. If the results are proven to be safe for the patient, then the documents will be ready for regulatory submission.

If a patient safety concern arises, then possible risk mitigation steps will have to be included in the manufacturing process, or the component may need to be replaced. If the patient safety concern still exists, then the characterization will need to be done by leachables testing followed by re-evaluation for patient safety. If the results are proven safe to the patient, then the documents would be ready for regulatory submission at that point. However, if after the re-evaluation the safety concern still cannot be resolved, then the component will need to be replaced to ensure patient safety.

 <665>, Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products. 

 <1665>, Characterization and Qualification of Plastic Components and Systems Used to Manufacture Pharmaceutical Drug Products and Biopharmaceutical Drug Substances and Products. 

 (Rockville, Md., 2024).
3. Shea, J. The Role of BioPhorum Extractables Data in the Effective Adoption of Single-Use Systems. Whitepaper, March 2022.
4. EMA. EMA/178637/2020—Rev.2. Summary on Compassionate Use (April 3, 2020).
5. FDA. VEKLURY Label. 

Ramesh Raju Mavuleti*, rrajum@gmail.com, is head of operations, Validation Services India. Subhasis Banerjee, PhD, is principal technical application expert, Bioprocessing APAC. Tathagata Ray is consultant, Global Strategy Deployment; all at Merck Life Sciences Pvt, Ltd. K. Vasantakumar Pai, PhD, is professor and chairman, PG Department of studies & research in Industrial Chemistry, Kuvempu University. K. Sreedhara Ranganath Pai, PhD, is professor, Department of Pharmacology, Manipal Academy of Higher Education. Somasundaram Gopalakrishnan is senior consultant, Global Biopharm Center of Excellence, Merck Pte. Ltd.

*To whom all correspondence should be addressed.

®
Vol. 37, No. 6
June 2024
Pages: 27–31

When referring to this article, please cite it as Mavuleti, R. R.; Pai, K. V.; Pai, K. S. R.; et al. Application of the Risk Evaluation Matrix as per 

<665> and <1665> for Evaluation of Leachables Risk from Single-Use Components Used in Biomanufacturing. 

Quality

Determining E&L risk from single-use components can be used to build the level of extractable profiling and PERLs.

Application of the Risk Evaluation Matrix as per USP <665> and <1665> for Evaluation of Leachables Risk from Single-Use Components Used in Biomanufacturing

Developing a robust and reliable strategy for analytical studies of a biosimilar in development is crucial for a successful regulatory pathway. The importance of biosimilar products is tied to the push for more affordable, accessible biological treatments. However, bringing a biosimilar to market has strict challenges because the biosimilar developer must have a large set of analytical similarity data compared to the innovator biologic. In addition, the biosimilar developer must also contend with demonstrating interchangeability (1).

Establishing biosimilar analytical studies

There are several key criteria to consider when establishing analytical studies for a biosimilar drug candidate to ensure its safety, efficacy, and similarity compared to the reference product, says Olaf Stamm, PhD, technical business development director, Biopharmaceutical Testing Solutions, Charles River Laboratories.

Among these criteria are comparability, biological activity, impurity assessment, and stability studies, Stamm notes. He explains that, for comparability, the biosimilar needs to demonstrate similarity to the reference product in terms of structure, function, purity, and potency. Analytical methods should thus be chosen or developed to compare critical quality attributes (CQAs) between the biosimilar and the reference product. “This involves assessing the physicochemical properties of the biosimilar, such as molecular weight, size, charge, and higher-order structure. Techniques [such as] mass spectrometry, chromatography, spectroscopy, and electrophoresis are commonly used for this purpose,” Stamm says.

The biological activity of the biosimilar should also be evaluated in analytical studies to ensure it matches the biological activity of the reference product. Analytical studies for determining biological activity may involve in-vitro assays, cell-based assays, or other relevant biological assays depending on the mechanism of action of the drug, Stamm remarks.

Identifying and quantifying impurities should include process-related impurities, degradation products, and contaminants. This is a crucial aspect of biosimilar development, Stamm notes. “Analytical methods should be sensitive enough to detect impurities at levels that are relevant to safety and efficacy,” he states. Stamm also points out that analytical methods should be established to identify and quantify residual host cell proteins to ensure they are within acceptable limits because their presence could impact safety and efficacy as well. “The evaluation of process-related impurities, such as residual solvents, reagents, and by-products, should be performed using appropriate analytical techniques,” he adds.

Meanwhile, stability studies are essential for assessing the biosimilar product’s shelf-life under various storage conditions. Analytical methods used for stability studies should monitor degradation pathways and stability-indicating parameters over time.

In biosimilars development, it is not enough to establish analytical evidence of the product’s biosimilarity to the reference product, but also its interchangeability. Luckily, establishing analytical studies for testing biosimilarity and interchangeability involves similar principles, Stamm explains. However, there are some key differences in the objectives and scope of these studies.

Stamm points out that the primary objective of analytical studies for demonstrating biosimilarity is to demonstrate that the proposed biosimilar is highly similar to the reference product in terms of quality, safety, and efficacy. “The goal is to show that any differences between the biosimilar and the reference product are not clinically meaningful,” he says.

In comparison, the objective of studies for interchangeability goes beyond demonstrating similarity. “[Interchangeability studies] aim to provide evidence that the biosimilar can be substituted for the reference product without any additional risk in terms of safety or diminished efficacy compared to using the reference product alone,” Stamm states.

Stamm further explains that interchangeability studies may involve additional clinical work. “Clinical studies for interchangeability are specifically designed to assess the impact of switching between the biosimilar and the reference product on safety and efficacy outcomes. These studies may include switching studies, pharmacokinetic (PK) and pharmacodynamic (PD) assessments, and immunogenicity evaluations in both naive and switch populations,” he says.

Both biosimilarity studies and interchangeability studies provide information that is crucial for regulatory authority evaluation. Stamm emphasizes that each type of study yields required data as explained in the following sections.

 According to Stamm, biosimilarity studies generate extensive analytical data comparing the biosimilar product’s CQAs with the CQAs of the reference biologic, including physicochemical characterization, biological activity assays, impurity profiling, and stability data.

 Meanwhile, clinical studies provide evidence of equivalent efficacy, safety, and immunogenicity between the biosimilar and the reference product in sensitive patient populations. These studies typically include PK/PD assessments as well as comparative clinical trials in relevant patient populations.

Immunogenicity data. Immunogenicity data—including an assessment of the development of anti-drug antibodies—is also a critical component of biosimilarity studies because these data help evaluate the potential risk of immunogenic reactions to the biosimilar.

 Interchangeability studies yield data on the safety and efficacy of switching between the biosimilar and the reference product, notes Stamm, who explains that these studies also include assessments of PK/PD, immunogenicity, and clinical outcomes following multiple switches between the two products.

 Data from interchangeability studies also provide insights into patient outcomes from the impact of switching medications, including any potential risks associated with switching between the biosimilar and the reference product.

Immunogenicity with switching. Immunogenicity of the biosimilar product is also assessed in patients who undergo repeated switching between the biosimilar and the reference product.

 Regulatory authorities require evidence demonstrating that switching between a biosimilar and its reference product neither compromises clinical outcomes nor poses additional risks to patient safety or efficacy compared to using the reference product alone.

“Overall, both biosimilarity and interchangeability studies generate comprehensive data sets that regulatory authorities evaluate to assess the similarity, safety, efficacy, and interchangeability of biosimilar products compared to their reference products. These studies are essential for the regulatory approval of biosimilars and for ensuring their safe and effective use in clinical practice,” Stamm concludes.

1. Mirasol, F. Moving Biosimilars Forward in a Hesitant Market. 

®
Vol. 37, No. 6
June 2024
Pages: 32–33

When referring to this article, please cite it as Mirasol, F. Reviewing the Importance of Biosimilarity and Interchangeability. 

Analytics

Biosimilar analytical assessments focus on demonstrating biosimilarity and interchangeability with the reference biologic.

BioPharm International, June 2024

Speeding Up Aseptic Processes

Addressing the Complexities of Media Formulation Development

Evaluating the Use of Continuous Chromatography

Innovations in Prefilled Biologics

Application of the Risk Evaluation Matrix as per USP <665> and <1665> for Evaluation of Leachables Risk from Single-Use Components Used in Biomanufacturing

Reviewing the Importance of Biosimilarity and Interchangeability

Publisher's Note: Automating Processes

Elements for a Sustainable Environmental Monitoring Program