Feliza Mirasol

When it comes to cleaning validation in the biopharmaceutical industry, different biopharmaceutical manufactures face similar challenges. Some common challenges include the ability to have reproducibility when manually cleaning small parts and when using non-clean-in-place (CIP) systems as well as having an effective design in which CIP systems are fully automated.

	Other challenges include a physical build-up of protein on equipment walls that have been cleaned by CIP. This build-up is a process that happens over time when there is no periodic scrub-down of the equipment, allowing for a layer of residue to accumulate in the inner surface of the vessel. In addition, failure of CIP systems over time due to blocked steam traps, poor maintenance, or irregular/erroneous replacement of a steam trap will pose problems for cleaning validation operations (1).

	Because cleaning validation is an important aspect in current good manufacturing practice (cGMP) manufacturing for monoclonal antibodies (mAbs) as well as other biologics, it is important to consider how current practices ensure that cleaning and sterilizing biomanufacturing equipment is conducted properly, and whether differences exist between cleaning and sterilizing equipment used in multimAb product versus single-mAb product manufacture in the same facility.

	To start with, no real differences exist between single-mAb and multimAb product manufacture in regard to cleaning and sanitation/sterilization, industry experts say. Cleaning protocols would be similar in both circumstances. However, it is important to note that there would be no concern with active carry-over in singleproduct facility as there would be in a multi-product facility.

	In the case of AstraZeneca’s Frederick Manufacturing Center in Frederick, MD, for example, the acceptance criteria for sterilization in place (SIP) is based on the European standard EN285 and is non-product specific, according to an AstraZeneca company official.

	“The acceptance criteria for cleaning are non-product specific because sampling assays and each [n]ew [p]roduct [i]ntroduction (NPI) are challenged on a small-scale (i.e., recovery/cleanability), and protein denaturation (i.e., degradation of proteins using in-house chemicals) needs to be demonstrated to verify that the facility is fit. Each program (cleaning/SIP) has on-going monitoring/maintenance program executed annually to verify systems remain in a validated state,”the AstraZeneca official states.

	“Cleaning protocols between a single-mAb and a multi-mAb manufacturing facility would assess the degraded product as an organic impurity in the drug substance of the next batch,” notes Paul Lopolito, senior manager, STERIS Technical Services. “In a multi-mAb product facility, the main difference is having to assess the cross-contamination risk of mAb product process contaminants with the quality attributes of a subsequent, different mAb drug product,” adds Beth Kroeger, manager, STERIS Technical Services.

	“A risk assessment should include an assessment of active degradation rinsability, toxicity, host cell proteins, media, cleaning agent residues, cleaning process capability, and bioburden and bacterial endotoxin,” notes Kroeger.

	In a biomanufacturing facility where multiple mAb therapeutics are produced, it is important to note what types of cleaning validation are necessary, and how often validation must be performed. The AstraZeneca official notes that cleaning validation maintenance is performed annually at the company’s Frederick facility using a matrix approach. “For each NPI, small-scale cleanability/recovery, protein denaturation, and large-scale cleaning verification (matrix approach) are required,” the official states. In addition, establishing if the product is more challenging to clean compared with previous products will determine the requirements for cleaning validation/verification.

	Lopolito points out that a risk assessment should be performed for incoming mAb drug-product toxicity, solubility, degradation product rinsability, media, and buffer constituents compared to the existing validated cleaning procedures for the entire process for the current products. “If this new mAb is not considered worst case, then at least one production run under normal process conditions for cleaning validation or verification should be executed. Provided no significant changes are made to the process, revalidation is generally not required. Continued process verification should be in place to trend quality attributes to establish cleaning process capability and performance control,” he explains. 

	Systems must be in place that help maintain a sterile environment for manufacturing. In terms of cleaning and SIP validation, all equipment, systems, and production parts used for manufacture should be cleaned with a validated cycle prior to use, the AstraZeneca official says. “Cleaning cycles demonstrate that equipment surfaces are clean and free of residues and ensure any leftover product is cleared,” the official states. 

	Kroeger notes, however, that no additional systems are required in a multimAb facility compared to a single-mAb facility “unless the facility is manufacturing products such as antibody-drug-conjugates (ADCs), due to the potential increased toxicity from the small-molecule portion of the ADC, or products other than mAbs/large molecule.”

	Procedural segregation should exist with separate area clearance between products and batches in a multi-mAb facility, Lopolito adds. “Obviously, packed columns and ultrafiltration membranes would be product dedicated, but equipment need not be dedicated once the resin or membranes are removed. Equipment change-over between products is recommended where elastomeric parts are discarded but is not necessary if cleaning validation testing verifies residue removal from the various materials of construction, or they are not considered to have a significant surface area to exceed the maximum allowed carry over limits,” he explains.

	In addition, other procedures, components, and systems that are important to have in place for cleaning validation include automation control, gowning, aseptic practices, changeover requirements, high-efficiency particulate air (HEPA) filtered air, closed systems used for manufacture, positive pressure, isolation valves, external equipment cleaning, room classifications/cleaning, and on-going quality control monitoring, the AstraZeneca official emphasizes.

	Meanwhile, the selection of cleaning agents requires certain criteria. Lopolito points out that cleaning agents should be selected based on performance, rinsability, analytical methods availability, low toxicity, lot traceability, and compatibility with materials of construction.

	Safety is the most important consideration because of toxicity risks, adds the AstraZeneca official. “Do not select cleaning agents that have significant cytotoxicity, allergenicity, or reproductive hazards,” the official asserts. 

	In a multi-mAb manufacturing scenario, residue type is also a key consideration when selecting a cleaning agent because it is necessary to ensure that the cleaning agent degrades the residue of each product, product-by-product, the AstraZeneca official says. “Typically, alkaline cleaning agents are used as they are bactericides and emulsifiers [microbial control and dissolving proteins]. Acidic cleaning agents may be used to remove salt deposits. Cleaning chemicals with surfactants and builders, that are compatible and safe, are preferred,” the official says.

	Innovations in cleaning/sterilization materials are also important to the process. According to Kroeger, there is at least one cleaning agent that is registered as a biocidal agent with disinfectant, biocidal, and biofilm remediation claims. “Detergent performance enhancements including maintenance derouging, antimicrobial effectiveness, and biodegradability are a few of those being developed to support manufacturers’ process optimization and sustainability goals,” she states.

	In addition, a risk-based approach to acknowledge potential failure modes and provide justification on current controls (detection) is in place. It is important to implement this risk-based approach while maintaining the validated state and regulatory requirements, the AstraZeneca official says.

	Biostatistical analysis may also be used to establish control limits (within predetermined acceptance criteria) used for continuous monitoring/trending.

Articles

Current practices for cleaning and sterilizing biomanufacturing equipment does not differ much between a multi-product versus single-product setting.

Cleaning Validation for Biologic Equipment

The biopharmaceutical industry has long been working to refine and improve the biomanufacturing process to improve monoclonal antibody (mAb) productivity. Some success has been achieved in upstream bioprocessing where cell titer has been greatly increased as the result of better cell-culture methods and improved bioreactor performance, as well as optimized nutrition in cell-culture media and supplements. Biomanufacturers, however, continue to seek new and improved ways to increase productivity, including innovations in bioprocessing.

	The biggest challenge in mAb production is improving mAb titer without impacting product quality, and this is dependent on the stability of the cell lines used as the source and the expression systems with which the mAbs are produced. In upstream processing, determining the duration of the cell-culture process, the relative low density of the cells, and/or accumulation of stress-generating byproducts such as lactate or ammonium are also critical challenges.

	In addition, there still remains quite of bit of work to be done on the upstream side to increase yields, including work needed to fully characterize all the materials going into upstream processes. Currently, three grams of API per liter is traditional for mAbs, but some biomanufacturers are claiming higher concentrations at 10 g/L and up to 50 g/L, which, from a scale-up perspective, sounds aggressive, notes Claudia Berrón, vice-president of global commercial development, biopharma, at Avantor, a manufacturer and distributor of products, services, and solutions to the life-sciences and advanced technologies industries.

	Depending on their characteristics, each raw material can react differently with the drug molecule, and many raw materials going into upstream cell-culture processes have not been fully characterized or optimized for the target molecule, Berrón explains. Thus, upstream optimization needs better characterization of all media components, and, more specifically, anything going into the cell-culture supplement with that particular molecule is crucial as people seek higher upstream productivity.

	Improving productivity has largely been a question of progress in three areas, adds Nigel Darby, advisor to the CEO, GE Healthcare Life Sciences. First, mAbs were expressed at low levels in early bioprocessing in the early 1990s, typically 10- to 20-fold lower product titers than today, which necessitated the use of large bioreactors to manufacture sufficient product quantities. Second, downstream purification was limited by chromatography resins with four to five-fold lower capacity than today, particularly in the Protein A capture step, which necessitated the use of large volumes of resin to bind the product and wide-diameter chromatography columns to cope with the large volumes of dilute, clarified cell-culture supernatant coming out of the process. Third, both aspects of low productivity required big complex factories to manufacture significant quantities of mAb, which required high capital investments, long construction times, and significant financial risks if products failed in clinical trials, Darby explains.

	To tackle these challenges, technology has fortunately been rapidly evolving to provide solutions. In the past 10 years, significant improvements in cell culture titers have been achieved, with 5–10 g/L being achieved and around five-fold improvement in Protein A resin capacity, Darby asserts. “The net effect of these improvements has considerable impact. Bioreactors have shrunk from 20,000-L volume to 2000 L, and two-meter chromatography columns are replaced with 50–80-cm columns … all to achieve the same production output,” he states.

	In addition, single-use technology is beginning to replace the fixed stainless-steel facility designs that were once prevalent, particularly large stainless-steel bioreactors are being replaced with much smaller single-use bioreactors. Today, the most advanced biomanufacturing facilities are running with smaller unit operations and reduced infrastructure because of single-use technology, which is driving significant reduction in capital investment and construction times. Furthermore, the financial risk profile is much more favorable with scaled-down, right-sized facilities using single-use technology, according to Darby.

	To meet the challenges faced today and efforts to improve mAb productivity, design and development engineers are employing several methods or approaches to resolve these issues. For example, at Agenus Inc., the company’s design engineers and scientists look into ways of developing efficient and high expressing vectors for cloning cell lines and sometimes may be sub-cloning for optimization. The company now has cell lines with excellent growth rates and high specific productivity for both fed-batch and perfusion cultures for several immunoglobulin G (IgG) isotypes, says Al Dadson, head of Global Biologics Manufacturing, Agenus, an immuno-oncology company specializing in antibodies, vaccines, cell therapies, and adjuvants for treating cancers.

	“Our development engineers and scientists are working on both upstream, downstream, and analytical/formulation development platform processes to increase aspects of robustness, scalability, and reproducibility to complement high expression systems to reduce COGS [cost of goods sold],” Dadson says.

	Meanwhile, at Avantor, the company has been focusing on dramatically improving materials characterization for upstream processing. The company is specifically working directly with mAb producers to find the right combination of cell-culture components to optimize upstream yields and has recently implemented new laboratories, one focused on mammalian upstream cell culture and another focused on microbial fermentation, at its facility in Bridgewater, NJ, according to Berrón.

	In downstream bioprocessing, Avantor has process engineers working on optimizing three parameters: yield, process time, and purity. “Depending on the target protein molecule and other factors, we can optimize various parameters such as resin chemistry, buffer type, and specific additives that can modulate separation performance to get higher yield or higher purity, or we can reduce the number of process chromatography steps. In addition, we can optimize buffer conditions and flow rate through the columns to decrease processing time,” says Dr. Nandu Deorkar, vice-president of research and development, Avantor.

	“One area where we are having success is more targeted ligands. Certain feedstock may contain closely related product impurities that may require multiple traditional chromatographic steps. It is a common challenge we see in recombinant protein, beyond traditional mAbs,” Deorkar further states. “To address this challenge, we have been engineering mixed-mode, multimode, and hydrophobic chromatography resins that help optimize the removal of these closely-related impurities. This method has mainly been applicable to any recombinant protein that is produced in a microbial fermentation process,” he adds.

	Process engineers and scientists are also developing refined processes, such as implementation of perfusion steps, adds Melanie Diefenbacher, PhD, scientific consultant, Genedata, a data intelligence company that offers an integrated enterprise workflow platform to streamline R&D workflows and improve productivity. To achieve this refinement, they try to take a broader, integrated view on the overall process from the development of the manufacturing cell line up to the downstream and formulation steps. Process engineers and scientists are also using miniaturized, automated approaches to study factors influencing manufacturing success. “Therefore, digitalization methods, such as artificial intelligence and deep learning, are becoming increasingly important to support these efforts,” Diefenbacher says. “Using cyber-physical systems, the Internet of things, and cognitive computing, supported by smart laboratories and integrated and scalable information technology (IT) systems, R&D operations, and manufacturing systems are becoming decentralized, and at the same time intelligent, flexible, and highly integrated.”

	For the most part, the biopharmaceutical industry still has some challenges with assimilating the advances made in recent years to improve mAb productivity. There remain many operations that have not yet started to use the best technologies that are available today, according to Darby. “Given the lifecycles of many processes, it can be difficult to assimilate new technology quickly because of the constraints of regulation and cost as well as risk of change,” he says.

	Darby points out, though, that progress in technology continues, nonetheless. “For example, in the challenging downstream area, we are moving to consider new higher productivity technologies in purification, such as the use of nanofiber products as an alternative to chromatography beads. On the other hand, there are things we can do better with the resources we have today, [such as] better use of manufacturing process and raw material data to optimize manufacturing output and quality.”

	Compared to other industries, biopharmaceuticals are at a relatively early stage, but given the complexity of biomanufacturing, there are likely rapid gains to be made simply by understanding, for example, the effects of raw material variation on productivity and better understanding of how to control and predict that, Darby asserts.

	Another area for potential bioprocessing productivity improvements is in downstream processing, where single-use technology can be combined with process chromatography columns, says Deorkar. “Using single-use systems has the potential to move from a batch-based approach to something approaching a connected continuous process. It will be possible to minimize the large storage tanks and use single-use systems to streamline how you collect the samples, how you load the samples, and so forth. There are single-use systems that make it much more of a continuous process and remove the time required to clean, dry, qualify, and validate sampling and storage systems between chromatographic steps,” he states.

	Single-use systems have the potential to improve productivity across the entire mAb production process, especially in terms of sampling, adds Berrón. “Drug producers have to sample for quality and process control purposes many, many times continuously. Typically, to effectively sample, you sample more than what is needed, and, in some cases, it isn’t possible to sample exactly what is needed.”

	The industry is using approaches like perfusion or some sort of continuous manufacturing to increase volumetric output and increase yield, in addition to developing high-producing cell lines, confirms Dadson. “Systems like the ATF [alternating tangential flow] and other tangential flow systems have been known to increase productivity by several fold. The use of PAT [process analytical technology] in platform facilities is another avenue being explored by industry,” he says.

	Dadson also notes that certain approaches in cell-line engineering are using Chinese hamster ovary (CHO), nonsecreting null (NS0), Sp2/0, HEK293, and PER.C6 cell lines as host systems, although more than 70% of industry today is using CHO cells. New approaches also include transgenic plant and animal systems for host cells.

	The Agenus West team in Berkeley, CA, for example, is working with Agenus’ cell line-development team in Cambridge, UK, and its Discovery/R&D teams in Lexington, MA, to ensure cell lines are developed with the required product quality attributes and commercial manufacturability. “The availability of desired cell lines for commercial production shortens manufacturing time for drug development. Coupled with this fully integrated approach, Agenus West thrives on the latest cutting-edge technology platforms in-house, making us self-reliant and giving us the advantage of manufacturing speed, cost efficiency, operational flexibility, and manufacturing technology transfer to commercial scale partners-all with desired product quality,” Dadson states.

	Approaches to improving mAb productivity have included increasing throughput and reducing cycle times using new automation equipment, says Diefenbacher. Another approach involves using new and refined bioprocesses that include the application of fully or at least partially continuous processes, such as upstream perfusion process steps. “In terms of newer innovation, the biggest push right now in productivity improvement is expected to come from the digitalization of the biopharma industry, which will improve process robustness and product quality and result in a more efficient use of resources,” Diefenbacher states.

	Future mAb production facilities are expected to see the implementation of a broad range of technologies, including the use of nanofiber technology in place of chromatography beads, increased automation and digitization of processes, and single-use systems. A few facilities are already exploring the use of some of these technologies today with many new facilities under construction, Darby observes. Meanwhile, innovations such as the nanofiber technology are still in development.

	“The smaller highly flexible facility with a high content of single-use technology seems to be the future for much of the industry. Instead of scaling up, many now talk of scaling-out-building capacity as needed by rapid construction of new small-scale facilities rather than taking the risk to build a single large facility,” Darby says.

	As the mAb production process continues to intensify, the lack of “downtime” that used to occur naturally as a consequence of the greater demands of cleaning and maintenance in standard infrastructure puts ever more pressure on manufacturing teams and extended supply chains to become more efficient, Darby also points out.

	“The burden of some activities will be transferred to suppliers, as we see with the implementation of single-use technology, and this is expected to increase. We are still undergoing a significant transition in implementation of single-use technology with more operator training required to ensure these technologies can be applied successfully, particularly as productivity increases,” he states.

	“Right now, the biggest push is expected to come from the digitalization of the biopharma industry, which will improve process robustness and product quality and result in more efficient use of resources,” Diefenbacher adds, noting that most big pharmaceutical companies today are already looking for ways to directly capture the huge amounts of data produced in R&D and manufacturing, such as cell-line selection, process optimization, or media development.

	“The implementation of a central data backbone for sharing all data and driving digitalization across all development groups to connect information from upstream, downstream, formulation, and analytics development units is essential for every biopharma organization,” she says.

	“We think there will be more use of single-use technology, although there will still be plenty of need for stainless-steel, fixed production systems,” Berrón interjects. Blockbuster drugs would still need stainless-steel production systems, she explains, because they have long production runs, and using stainless-steel is the most efficient way to produce a blockbuster drug. For faster change and faster turnarounds where smaller-volume drugs are involved, single-use technology continuously gains ground in this space.

	“We also see greater investment and greater use of advanced data tools, and this is something that Avantor is making significant investments in-digital tools for process optimization,” adds Deorkar. Compared to other industries, even small molecule, the biopharmaceutical industry is lagging in terms of how data is captured, what kind of data is captured, and how it is mined and used productively, he says.

	There is also a push for future bioprocess facilities to build efficient continuous processing facilities, Dadson states. The push for facilities that use high-expression cell lines in combination with end-to-end single-use systems to build smaller, cost-effective footprint facilities versus conventional stainless-steel facilities has become an industry focus. “One drawback is the maximum size of single-use systems (2 kL–3 kL) to handle high product demand, especially in oncology, versus >20 kL stainless-steel facilities,” he says.

	Finally, the increase in automation to replace low-value activities, such as buffer management, and an increase in real-time data analytics in the management of processes will also require whole new skill sets in the workforce, Darby points out. “Time pressures will likely drive towards more analytical work being performed at or on-line during manufacturing, with paperless workflows. These innovations are just emerging in the most advanced operations but will require changing approaches from plant operators.”


	Vol. 43, No. 5
	May 2019
	Pages: 16–22

	When referring to this article, please cite it as F. Mirasol, “The Search for Bioprocess Productivity Improvement,” 

Large biopharmas, emerging biotechs, and CMOs are looking for novel ways to improve the productivity of biologics in a rapidly evolving biotherapeutics market.

The Search for Bioprocess Productivity Improvement

	Biostatistical analysis may also be used to establish control limits (within predetermined acceptance criteria) used for continuous monitoring/trending. 


	Vol. 43, No. 5
	May 2019
	Pages: 50–52

	When referring to this article, please cite it as F. Mirasol, “Managing Biologic Equipment Cleaning,” 

This article takes a look at current practices for cleaning and sterilizing biomanufacturing equipment used in a multi-product versus single-product setting.

Managing Biologic Equipment Cleaning

Biopharmaceutical manufacturers have embraced the use of single-use technology upstream, but are now increasing its use in downstream processing (DSP) as well. “There are very few DSP unit operations that are not using single-use systems or components,” notes Donald Young, senior global product manager, Thermo Fisher Scientific, BioProduction Division. “Clarification via depth filtration or tangential flow membrane filtration (TFF) is just one example.”

	As this trend continues, the concept of an end-to-end single-use manufacturing facility has moved from the world of theory squarely into reality. However, downstream single-use systems are still evolving, as vendors work to improve performance in critical areas. This article summarizes some of the benefits and the challenges of employing single-use technology downstream.

	Reduction in bioprocessing time is one advantage that single-use systems offer. As Young points out, re-usable systems can typically require more than 12 hours from set-up to teardown, while single-use TFF systems require half that time. 

	Another area where single-use is gaining ground downstream is in buffer preparation, storage, and dispense. “Single-use mixers, coupled with single-use bioprocess containers (BPC), can increase a DSP suite’s annual throughput, because they offer turnaround times of around a day or less without the need for clean- or steam-in-place operations. Reusable systems can require three or more days to clean and sterilize,” Young explains. 

	Prefilled BPCs have only made the single-use option more popular, he says, noting that BPCs prefilled with water for injection or purification buffers can be delivered to customers, eliminating the need for them to perform these functions inhouse. These systems can be supplied in volumes ranging from pre-clinical and clinical scale to volumes large enough to satisfy current good manufacturing practice (cGMP)-scale production, he says.

	“These large-volume-liquid BPCs are used as the primary source of sterile process liquids or as a second source/back-up supply to reduce demand on the customer’s in-house capabilities and capacities,” Young states.

	Many disposable chromatography columns meet cGMP and industry standards, and also offer reproducibility, scalability, speed, ease-of-use, and operational safety. However, chromatography has been the last purification stage to move into disposable equipment. Although its use is growing downstream, it is still limited. In addition, the industry’s adoption of pre-packed, limited-use chromatography has been slow, even though the technology’s benefits have been demonstrated in the early stages of clinical manufacturing (1).

	Among the common problems that single-use technology can help address is column packing. Column packing requires a stable bed with good flow properties, and, today, automatic packing solutions make packing the columns much more reliable than in the past. Bioburden can still result, however, and its source can vary-usually it is introduced before or during packing. Thus, good sanitation methods and familiarity with the most likely microbial contaminant species present in the immediate environment are needed to address bioburden. Using pre-sanitized single-use columns and flow kits is one way to minimize the risk of bioburden (2).

	Single-use technology implementation in chromatography can greatly reduce cycle times, lower operating costs and energy consumption, and generate less waste, according to Young, who points out that DSP can be a process bottleneck because of a lack of single-use chromatography options or because of the expense, time, and labor of using existing chromatography components as single-use components.

	With a facility set up for single-use system operations, facility design can be simplified and can be executed in about half the time needed to build a traditional facility with reusable systems, Young emphasizes. “Start-up/licensure times can be cut compared to traditional facilities using stainless-steel-only equipment and systems,” he states. In addition, construction costs are lower mostly due to reduced need for clean steam systems that can account for much of a production facility’s water needs, he adds.

	“Single-use systems are an attractive option for emerging markets, not only in chromatography but all along the production workflow,” says Young, who sees this as the technology’s most important contribution today. 

	Despite wider adoption of single-use technologies, downstream bioprocessing, particularly chromatography, continues to a pose challenges. Available technology may not always be adequate to meeting a specific facility’s needs. For example, not all downstream bioprocessing steps are available with single-use technology, and those that are available may be limited by performance, scalability, and cost (1).

	For example, one unmet user need is the lack of affinity and ion-exchange chromatography beads that are sufficiently cost-effective to be classified as single-use. These process steps continue to be the biggest process bottleneck in the DSP workflow, and represent a huge opportunity for suppliers, Young remarks.

	Another area where single-use technology might be improved downstream is in combining it with process chromatography columns, according to Dr. Nandu Deorkar, vice-president of research and development at Avantor, a manufacturer and distributor of products, services, and solutions to the life-sciences and advanced technologies industries.

	“[Adopting] single-use systems gives manufacturers the potential to move from a batch-based approach to something approaching a connected continuous process. It will be possible to minimize the number of large storage tanks required, and incorporate single-use systems to streamline sample collection and loading, and other operations. There are single-use systems that make it much more of a continuous process, and remove the time required to clean, dry, qualify, and validate sampling and storage systems between chromatographic steps,” Deorkar states.

	Another unmet need for biopharmaceutical manufacturers is technology that reduces the cost of resins (3), to allow for greater use of single-use systems to address the resin fouling that occurs from repeated use of resin during chromatography. Manufacturers are unlikely to dispose of costly resins after one pass, unless costs are significantly reduced to make this practical.

	Because chromatography resins make up a large portion of the costs in downstream bioprocessing, switching a polishing step from resin chromatography to a single-use membrane column can significantly reduce expenses and is one alternative when looking to improve DSP. In a study (4) that used modeling software to evaluate commercial monoclonal antibody (mAb) production, it was shown that the unit operation cost was 19% to 33% less for a membrane process used on 1000-L and 5000-L scales bioreactor volumes and various mAb titres. In addition, buffer volume was decreased by up to 55%.

	Suppliers of single-use downstream chromatography systems continue to improve the options available to manufacturers, and to examine ways to reduce the number of processing steps required for biomanufacturing, by linking steps to improve process efficiencies. For example, the ability to use concentrated buffer components and mix them in-line under controlled conditions (i.e., in-line conditioning) has simplified chromatographic operations (2).

	“Single-use systems have the potential to improve productivity across the entire mAb production process, especially in terms of sampling,” adds Claudia Berrón, vice-president of global commercial development, biopharma, Avantor. “Drug producers have to sample for quality and process control purposes many, many times continuously. Typically, to effectively sample, you sample more than what is needed, and, in some cases, it isn’t possible to sample exactly what is needed. Every time you take a sample out, you’re taking valuable material out of the process.”

	For its part, Avantor has been creating single-use systems that incorporate design features that make it easy to customize sampling systems to extract only the amount of material required to perform a test or document process results. “These designs are also much easier for plant personnel to use, helping reduce the time and labor costs associated with sampling,” Berrón states.

27 (6) 34–36 (2014).
	2. C. Hroncich, 

29 (9) 28–33 (2016).
	3. R. Jacquemart et al., 

Computational and Structural Biotechnology Journal

14, 309–318 (2016).
	4. A. Xenopoulos, 


	Vol. 43, No. 5
	May 2019
	Pages: 40–42

	When referring to this article, please cite it as F. Mirasol, “Single-Use in Downstream Chromatography: Benefit or Hindrance?” 

Single-use technology is gaining ground in downstream bioprocessing, but challenges stall further adoption.

Single-Use in Downstream Chromatography: Benefit or Hindrance?

	Editor's Note: This article was published in 

In the European Union (EU), the landscape for biosimilar medicines continues to be robust, with 16 products representing six biosimilar molecules granted marketing authorization by the European Commission (EC) in 2018. This number is consistent with the biosimilar marketing authorizations granted by the EC in 2017 (1).

	Dominating the marketing authorizations was Novartis’ Sandoz, which garnered five biosimilar product approvals, or 31% of total approvals. Pharma major Mylan followed close behind with four biosimilar product approvals, or 25% of total approvals. Amgen also won multiple marketing approvals, and Pfizer received one. Other companies that received marketing authorization for biosimilars include Accord Healthcare Limited, Celltron, Cinfa Biotech, and ERA Consulting (see 

	Between Sandoz, Mylan, Amgen, and Pfizer, pharma majors accounted for 75%, or 12, of total 2018 EU marketing approvals for biosimilar products. The six biosimilar molecules these products represent are adalimumab, bevacizumab, infliximab, insulin glargine, pegfilgrastim, and trastuzumab.

	Sandoz’s five marketing approvals include Zessly (infliximab), Ziextenzo (pegfilgrastim), and three adalimumab biosimilars: Halimatoz, Hefiya, and Hyrimoz. Zessly is a biosimilar referencing Remicade (infliximab), Johnson & Johnson’s top-selling drug with US$5.3 billion (€4.6 billion) in 2018 sales (2). In the EU, Zessly is indicated for treating rheumatoid arthritis, Crohn’s disease, ulcerative colitis, ankylosing spondylitis, psoriasis, and psoriatic arthritis.

	Sandoz’s three biosimilar adalimumab products-Halimatoz, Hefiya, and Hyrimoz-reference AbbVie’s Humira (adalimumab), which is AbbVie’s top-selling drug with nearly US$20 billion (€17 billion) in 2018 sales (3). Hefiya is indicated for plaque psoriasis, psoriatic arthritis, axial spondyloarthritis, polyarticular juvenile idiopathic arthritis, acne, and uveitis; Halimatoz is indicated for the same conditions plus active enthesitis-related arthritis, and ulcerative colitis; and Hyrimoz is indicated for all the same conditions as Halimatoz as well as Crohn’s disease.

	Ziextenzo, indicated for treating neutropenia, is a biosimilar referencing Amgen’s Neulasta (pegfilgrastim), also a blockbuster drug with approximately US$4.5 billion (€3.9 billion) sales in 2018 (4).

	Similarly endowed, Mylan also received marketing approvals for biosimilars referencing blockbuster innovator biologics. Mylan’s approvals include Semglee (insulin glargine), Hulio (adalimumab), Fulphila (pegfilgrastim), and Ogivri (trastuzumab). Semglee references Sanofi’s Lantus, which earned €2.7 billion (US$3.1 billion) in the first nine months of 2018 (full-year 2018 sales data not available at time of this article’s publication) (5). Ogivri references Roche’s Herceptin, which garnered CHF 5.3 billion (€4.6 billion, US$5.3 billion) in the first nine months of 2018 (full-year 2018 sales data not available at time of this article’s publication) (6). It is indicated for breast and stomach cancer.

	As with Sandoz’s biosimilars, Mylan’s Hulio references AbbVie’s Humira and is indicated for treating plaque psoriasis, psoriatic arthritis, rheumatoid arthritis, axial spondyloarthritis, polyarticular juvenile idiopathic arthritis, active enthesitis-related arthritis, Crohn’s disease, ulcerative colitis, acne, and uveitis. Mylan’s Fulphila references Amgen’s Neulasta and is indicated for neutropenia.

	With two biosimilar marketing approvals in 2018, Amgen’s products include Mvasi (bevacizumab) and Kanjinti (trastuzumab). Mvasi references Roches’ Avastin (bevacizumab), another blockbuster biologic with CHF 5.1 billion (€4.4 billion, US$5.1 billion) in the first nine months of 2018 (6). Mvasi is indicated for treating colon, breast, lung, kidney, ovarian, and cervical cancer. Kanjinti is another biosimilar referencing Roche’s Herceptin and is indicated for breast and stomach cancer.

	Also stepping up competition in oncology is Pfizer’s Trazimera (trastuzumab), another biosimilar referencing Herceptin. Pfizer’s product is also indicated for treating breast and stomach cancer.

	The other companies accounting for the rest of the biosimilar marketing approvals in the EU last year were Accord, an India-based global supplier of generic and speciality pharmaceuticals, Celltron, a South Korea-based biopharmaceutical company, Cinfa Biotech, a Spain-based biopharmaceutical company specializing in biosimilars, and ERA Consulting, a German-based consulting firm that specializes in product development and regulatory affairs.

	Accord, Cinfa Biotech, and ERA Consulting all received EU approval for their respective pegfilgrastim biosimilars, referencing Neulasta. Accord’s product is Pelgraz; Cinfa Biotech’s product is Pelmeg; and ERA Consulting’s product is Udenyca. All are indicated for treating neutropenia. Celltrion received approval for Herzuma (trastuzumab), also referencing Herceptin and indicated for breast and stomach cancer.

www.ema.europa.eu/en/search/search?sort=field_ema_med_market_auth_date&order=desc&search_api_views_fulltext=biosimilar

, accessed 23 Jan. 2019.
	2. Johnson & Johnson, “

Johnson & Johnson Reports 2018 Fourth-Quarter Results

,” Press Release, 22 Jan. 2019.
	3. AbbVie, “

AbbVie Reports Full-Year and Fourth-Quarter 2018 Financial Results

,” Press Release, 25 Jan. 2019.
	4. Amgen, “

Amgen Reports Fourth Quarter And Full Year 2018 Financial Results

,” Press Release, 29 Jan. 2019.
	5. Sanofi, “

Sanofi Q3 2018 Performance Confirms Return to Growth

,” Press Release, 31 Oct. 2018.
	6. Roche, “

Roche Reports Very Strong Growth in the First Nine Months of 2018


	Vol. 31, No. 2
	February 2019
	Page: 46–47

	When referring to this article, please cite it as F. Mirasol, “Roundup of European Biosimilars Approved in 2018," 

The market for biosimilars in Europe remains consistently robust, with 16 new biosimilars approved in 2018.

Roundup of European Biosimilars Approved in 2018

Client–vendor relationships require a level of trust, open communication, and reliable deliverables. Pharmaceutical Technology spoke with Chris Mooney, senior quality assurance technical specialist at Fujifilm Diosynth Biotechnologies, a contract development and manufacturing organization (CDMO) specializing in cell culture, fermentation, and viral vectors; and Knut Niss, chief technology officer of Mustang Bio, a US-based clinical-stage biopharmaceutical specializing in chimeric-antigen-receptor-engineered T cell (CAR T) immunotherapies and gene therapies, to discuss current best practices that help ensure that quality control is maintained during the course of the client–vendor relationship. Mooney gives his views from the perspective of a CDMO dealing with materials suppliers, and Niss discusses his experience from the perspective of a biopharma company working with contract service providers as well as material suppliers.

 What criteria are sought after in a vendor/service provider?

 Here at Fujifilm Diosynth Biotechnologies, we categorize our raw materials and components. The category of the material will determine the supplier requirements. The quality tools we primarily utilize are onsite supplier audits and audit questionnaires, plus supplier quality agreements. In order to become approved, suppliers must meet our acceptance criteria. Again, the material category will determine the frequency at which each supplier is re-evaluated.

 The number one criterion for us in vendor selection is quality. We expect vendors to provide a quality system that is up to our own standards. While we are only in early stages of development, we realize that having a strong quality system is critical for an expedited path through development. We do not want to find ourselves in a situation where we have exciting clinical data with great promises for patients and then have to slow down development in order to develop appropriate quality management at the vendor. This includes the ability of the vendor to safely provide materials/services on time.

 What systems or best-practice procedures are typically in place to ensure that there is quality control/quality oversight between vendors and their clients?

 A key aspect is first to build a relationship between our quality assurance (QA) and the vendor’s QA organization. Single points of contact are critical here as those assure timely and proper communication between the organizations. Procedurally, a well-designed vendor questionnaire and ultimately an audit assure a good relationship. Generally, the theme should always be: trust is good, control is better!

 All incoming materials used in cGMP manufacturing go through an evaluation, including inspections, or, in the case of raw materials, are tested. If an incoming material fails to meet our specifications, the material is rejected and a supplier complaint is initiated requesting the supplier to conduct an internal investigation.

	In addition to incoming material evaluations, supplier change notifications are also monitored. It’s important that agreements are in place with your suppliers to provide change notifications. QA initially evaluates supplier change notifications. The significance of the change will determine if a cross-functional team is required to evaluate potential impact to product quality.

 What tips do you have for keeping lines of communication open between vendor and client, and what is the impact of level of communication on the level of trust between vendor and client?

 Typically, supplier and client communications occur through our supply chain and procurement departments. We continuously look for ways to improve communications. Several of our larger suppliers have come onsite to provide details about the products and services they provide. Whenever you have an opportunity to provide your suppliers with an overview of your company, take it. It is important that suppliers understand the materials they provide are used to manufacture drugs for human use.

 The most important thing is to have single point of contact on both ends. If too many parties are involved, confusion and miscommunication set in, resulting in unnecessary delays. In this context, it is important that a backup is identified as well to assure an uninterrupted process in case the appointed contact is on vacation or otherwise out of the office.

 How do quality assurance practices differ when dealing with one-off vendor relationships versus longterm vendor relationships?

 For one-off suppliers, we generally apply the same rules. However, with long-term vendors, we like to establish a supply agreement that includes a quality agreement. With that, both sides know what to expect and can plan accordingly. Generally, we try to avoid one-offs as much as we can, unless there is a very compelling reason. For this, we follow ‘you get what you pay for’. In other words, cheap is not the only decision point, quality is.

 No difference from a quality perspective. All suppliers providing us materials used in cGMP manufacturing must meet the same supplier approval requirements.

	When referring to this article, please cite it as F. Mirasol, “Ensuring Quality Control in Vendor Relationships," 

A look at some best practices to ensure that quality control is maintained in the client–vendor relationship.