Feliza Mirasol

Because the raw materials used in biologic drug development can impact drug interaction and patient safety, the testing of raw materials is necessary to ensure quality. 

spoke with Roisin Browne, site manager at Almac Sciences’ analytical services site in Athlone, Ireland; Andrew Bulpin, head of Process Solutions at MilliporeSigma; and Erik Foehr, PhD, vice-president, Analytical Sciences, at Pacific BioLabs to discuss best practices for analytical testing techniques and instrumentation used for raw materials testing.

Pharmaceutical Technology’s Bio/Pharma Laboratory Best Practices 2018 

Bio/Pharma Laboratory Best Practices 2018 

 
	Vol. 42
	November 2018
	Pages: 38–41

	When referring to this article, please cite it as F. Mirasol, "Analytical Best Practices for Raw Materials Testing," 

Pharmaceutical Technology Bio/Pharma Laboratory Best Practices 

Articles

The testing of raw materials is essential as raw material quality determines the outcome of biologic product quality.

Analytical Best Practices for Raw Materials Testing

The increasing complexity of biologic drugs in development is in part driving the need for innovative delivery mechanisms. Innovation in syringe technology is one example of how drugmakers strive to address the challenges of administering these newer therapeutics.

	The rapid growth of newer and targeted molecular entities, such as monoclonal antibodies (mAbs), interferons, peptides, vaccines, and ophthalmics; the increasing prevalence of chronic diseases and cancers; and the growth of biosimilars are driving the growth of prefilled syringes. Another factor that contributes to this growth is the need for improved safety and ease of administration for patients and healthcare workers. Prefilled syringes are known to be easy to use and to need only a few administrative steps, which contributes to improved safety (1).

, Nicolas Eon, global product manager of syriQ at SCHOTT, and Claudia Roth, PhD, vice-president Innovation Management at Vetter, discussed key challenges, required standards, and the potential future impact of biologic drug development on the prefilled syringes market.

What are the main challenges or deficits in prefilled syringe technology today, specifically challenges meeting the needs of biologic drugs?

Over two-thirds of the drugs in the development pipeline of pharmaceutical companies are biologics. These drugs offer new treatment options for some of the hardest-to-treat diseases, such as cancer, and chronic diseases. Yet their highly complex molecule structures are more sensitive to temperature and environmental conditions and are prone to interactions with the pharmaceutical container. Hence, biologics require innovative packaging to ensure drug stability over the entire shelf life.

	With prefilled glass syringes, the drug comes into contact with up to seven materials, with each presenting a potential risk of interaction. Subsequently, it is of high importance to reduce this risk by using components made of high quality as well as optimized processes. For example, when it comes to gluing the needle, a process optimization of the gluing step can reduce drug/container interactions by minimizing the quantity of residuals glue monomer.

	Moreover, biologic drugs tend to be highly viscous, which means that more force is needed during administration. In addition, over 75% of biologics are used in combination with an auto-injector for self-administration. Subsequently, the prefilled syringe must meet highest accurate dimensions as well as ensure that the gliding performance remains consistent from one injection to the next. The mechanical strength of the glass container must also withstand the pressure applied during the administration process. 

What specifications are required for packaging biologic drugs in prefilled syringes?

As is often the case, creating easy-to-apply systems means a more complicated production process at the outset. As we know, a prefilled syringe system is a complex construct because it has many individual components. That is why it is critical to achieve the correct individual conjunction and interaction of its parts in order to enable both functionality and usability. For example, when administering a drug with a syringe, you need to have a plunger rod that operates in concert with the other components, especially the glass barrel. To achieve this, a slight coating of lubricant is placed on the inner surface of the glass barrel. Therefore, a key challenge is to define the appropriate coating process and to control the correct amount of lubricant needed to enable the best possible movement of the plunger rod. This must be achieved in a manner that avoids any form of interaction between the lubricant and the drug substance.

	In the biologics sector where the active ingredients of compounds are large and very complex molecules, their manufacture is equally complex and challenging. Thus, a vast degree of experience and a substantial level of process expertise is required. Biologics react with far greater sensitivity to environmental influences such as heat or light than other substances do. That is why it is important to design, develop, and implement the correct product-related production processes. For example, using a suitable dosing method and proper pumps in the filling process is critical.

	Another example is the fact that biologics can react negatively to the chemical composition of the stopper within the syringe. For manufacturers, this means it is important to have significant knowledge of packaging materials and components.

The Parenteral Drug Association’s (PDA) Technical Report (TR) No. 73, 

Prefilled Syringe User Requirements for Biotechnology Applications 

(2), published in 2015, can be used as a baseline to design syringes for biological drugs. Depending on the drug and the intended use, the detailed specification must be agreed upon between the PharmTech a company and the prefilled syringe supplier.

What innovations in prefilled syringes are currently on the market, or in development, to address the challenges that biologic drugs face?

SCHOTT recently launched syriQ BioPure, prefillable glass syringes that are specifically designed for self-administered biologics. These syringes ensure drug stability during shelf life and ease-of-administration for patients. The syringes are made of highly inert Fiolax borosilicate glass and are manufactured under improved processes to lower tungsten oxide and adhesive residuals. As more and more biologics are administered in combination with a device, the syringes work with leading safety and auto-injector devices, thus enabling patients to move from hospital to homecare. A uniform silicone layer ensures that the flow rate of the injection process is as smooth as possible, thus enhancing the patient experience.

In our opinion, despite all the innovative activities being considered today, quality will always be the leading criteria of achievement. After all, we are in an industry where patients literally get injected with the products we manufacture for pharma and biotech companies. It is for our customer, as well as the patient, that we make every effort to be as innovative as possible while striving to achieve the highest-possible manufacturing and process quality on a day-by-day basis.

	Another central target is achieving compatibility between the active ingredients of a drug and its method of administration. That means it has to be as ‘end-user friendly’ as possible. It is a well-accepted principal that the higher the usability of a drug-delivery system, the greater the acceptance among patients and caregivers. The reasons for this include the homecare sector where a patient-friendly system fits well and contributes to cost avoidance and timeintensive visit to doctors’ offices and hospitals. Also, patient adherence itself is strongly dependent on the convenience level of the drug-delivery system. 

	To put it simply, whatever protects or increases the safety of a drug-delivery system while increasing its usability, is an enhancement and an innovative advance for the market and, ultimately, for the patient.

What trends in biologic drug development do you think will have an impact on the future development of prefilled syringe technology?

One trend we see is the higher complexity of biologics as well as the higher sensitivity of the drugs. This results in the need for a superior extractables and leachables profile of the pharmaceutical primary packaging. More specifically, this means that the amount of tungsten and glue residual must be minimized, and the selected rubber formulations carefully considered. In addition, the options for lubrication, such as the use of silicone, must be evaluated.

	Another trend is the move from hospital to homecare to allow patients to self-administer the drugs in an environment where they feel most comfortable. To ensure that this is as easy and safe for the patient as possible, the self-injection is handled in combination with a device, such as auto-injectors. For the development of prefilled syringes this means that the container must be compatible with a number of devices and meet the specific requirements and dimensions.

	The rise in personalized medicine, including biologics, targeting small target groups also effects the development of prefilled syringes. Due to the smaller batch sizes that are required, a new supply chain concept and a flexible platform approach are needed. This flexibility can be introduced early on in the drug development process by using standard secondary packaging for the primary packaging. For example, SCHOTT recently launched its iQ platform concept in which syringes, including syriQ Bio-Pure, are packaged in a standardized tub, which simplifies the changeover from, for example, vials to prefilled syringes to run on one filling line. Subsequently, the syringe development needs to fit in with a broader picture in order to meet market demands.

	Lastly, the industry is confronted with more stringent regulations, which require the packaging to provide ‘design history file ready’ documentation and be ‘essentially free of particle’. SCHOTT continuously improves its process and online inspection technology to ensure compliance with current and future regulatory requirements. 

41 (10) 32–35 (2017).
	2. PDA, TR No. 73, 

Prefilled Syringe User Requirements for Biotechnology Applications


	Vol. 42, No. 11
	November 2018
	Pages: 28–29

	When referring to this article, please cite it as F. Mirasol, “Innovations in Prefilled Syringes for Biologic Drugs” 

As biologic drugs grow increasingly complex, drug delivery mechanisms such as prefilled syringes are being adapted to meet the challenges.

Innovations in Prefilled Syringes for Biologic Drugs

Although Chinese hamster ovary (CHO) cells are the standard in antibody-based drug production, there are limits to their use (1). Market pressures to develop more affordable biologic drugs quickly are also driving the need for optimizing and/or updating current methods of protein expression. Innovations in cell bank sources are attempting to address the needs in biopharma manufacturing for which a CHO cell line may be insufficient.

, Nicole Faust, CEO and chief scientific officer of Cevec Pharmaceuticals, and Mark Emalfarb, CEO of Dyadic International, spoke about the cell-line innovations their respective companies have taken on to bridge that gap in biopharmaceutical manufacturing. Cevec is using a human cellbased expression system, and Dyadic uses a fungus, 

, nicknamed C1, expression system that it has been developing for decades.

 What about your innovative cell line source differentiates it from the industry standard CHO cells?

 In contrast to the Chinese hamster ovary-derived cells, our Cevec amniocyte production (CAP) cell lines are of human origin. This has several advantages when producing recombinant proteins: the cells only add human post-translational modifications, thus avoiding any immunogenic structures, like non-human sugar structures attached to the protein. Moreover, any residual host cell proteins in the recombinantly produced product will be of human origin. The presence of non-human host cell proteins is a concern, in particular since there have been some cases (e.g., CHO derived blood coagulation factor IX) in which residual CHO host cell proteins have caused an immune response in the recipient. 

	A second differentiation criterion to CHO cells is the ability to produce complex glycosylated recombinant proteins. While the CHO-platform has been continuously optimized for the production of antibodies, CAP cells are particularly well-suited to express more complex proteins, such as, for example, blood coagulation factors or high molecular weight molecules. We have further engineered the CAP-Go platform to generate specific cell lines allowing the production of glycoproteins with tailor-made glycosylation patterns.

 CHO cells have a 20-hour doubling time, whereas our Eukaryota-engineered C1 cell line has a doubling time of only two hours, two times to 10 times higher productivity, and a production cycle of one-third to one-half the fermentation time with one-tenth the media cost compared to CHO. In addition, with our cell line, there is no need for viral inactivation. C1 is a better cell line than CHO because it is a natural enzyme (protein) producer that is more stable and can tolerate a wide range of growth conditions. In addition, C1 can express, secrete, and produce proteins such as antibodies at high levels (i.e., 2.4 grams per liter per day) and can easily grow in fermenters. In addition, C1 has a more stable genome than CHO since it is smaller and is haploidic. Therefore, it inherently has the potential benefits for becoming a successful protein production platform for biologics. In comparison, the CHO cell line was developed for many years before it became suitable as an industrial production platform.

 How and why did you choose this source from which to develop a technology platform to produce biotherapeutics?

 Fungi in general, and C1 in particular, have a natural efficient system for protein expression. C1 however, also has the following benefits:

		Low viscosity, since the morphology was changed to propagules

		High productivity that was generated by random mutagenesis and direct genetic manipulation

		Grows on simple defined media on various carbon sources

		Can tolerant high glucose concentration

		Easy scale up (was scaled up and used commercially to produce enzymes at greater than 100m

		Advanced genetic tools (efficient transformation).

 When developing the CAP platform, we were pursuing two goals: establishing a cell line from human origin that was not derived from tumors or from aborted embryos like the then-existing human cell lines and ensuring that the cells` histories were carefully documented. For this reason, we chose amniocytes (i.e., fetal cells obtained for prenatal diagnostics) as the primary source of cells. They were immortalized using defined DNA segments of the Ad5 genome, and every single step of generating the cell lines was documented in detail. Thus, the full history of the cell line is known as are all materials the cells have been in contact with. All this is included in a biologics master file deposited with FDA, which our clients can reference.

 What are the media and/or supplement requirements for growing and maintaining the population of these cells? Do these requirements differ from that used to grow and maintain CHO cells?

 The cells grow as singlecell suspensions to very high cell densities in serum-free, chemically defined media. The cells’ requirements are very similar to other suspension cell lines growing in serum-free cultures. For optimal process yields, fine-tuning and adjustment to the specific metabolic needs of the CAP cells are beneficial.

	One remarkable feature of the CAP cells is that they are robust in their growth behavior. For example, changes in media composition do not appear to result in changes to the pattern of posttranslational modifications.

 This is a complete synthetic and defined media that is based on glucose as carbon source, salts to provide macro- and micro elements, with a few free amino acids to supply organic nitrogen. It is very simple, robust, and low-cost media. Not many other organisms can grow and produce so well on such simple media.

	The control of the fermentation is also robust, based on a simple fed-batch strategy, requiring just control standard variables such as pH, dissolved oxygen, or foaming, which ensure scalability and provide versatility to the bio-manufacturing process. Additionally, the system is energy efficient and requires only little oxygen. 

 Is there a need for newer/innovative media and supplements in the market for emerging cell sources, such as yours?

 Since we are using simple defined already low-cost media that is based on glucose, we don’t think that there is a need for newer/innovative media and supplements in the market.

 For producer cell lines like CHO or CAP-Go, there are already excellent media available. However, when you produce very diverse products like we do with the CAP-Go platform, ranging from small growth factors to highly complex multimeric proteins, it becomes apparent that the optimal media/supplement composition depends very strongly on the particular product.

	With a growing number of complex recombinant proteins being manufactured, it would be very desirable to have ‘media optimization kits’ to be used to establish the optimal media composition during process development with the option to then obtain exactly this composition in GMP quality from the media manufacturer.


	Vol. 42, No. 11
	November 2018
	Pages: 40–41

	When referring to this article, please cite it as F. Mirasol, “Media Requirements for Different Cell Sources” 

This article will explore the requirements for media and supplements needed to maintain newer cell lines, such as those based on human cells and fungal cells.

Media Requirements for Different Cell Sources

This article was published in Pharmaceutical Technology Europe, Volume 30, Issue 8, August 2018

	Modular manufacturing is being increasingly used in the biopharmaceutical industry where flexibility and low-cost manufacturing systems are sought out. The design of a modular system in biopharmaceutical and pharmaceutical processing can involve breaking down a manufacturing facility into smaller functional building blocks, also known as modules (1).

	The term modular manufacturing is used throughout the biopharmaceutical industry in reference to modular facilities, modular processing equipment, or modular automation platforms, according to Joe Makowiecki, Enterprise Solution architect at GE Healthcare Life Sciences.

	“Any of these can fit the definition in that they involve modules as the basis of design or construction. Modular is particularly attractive for its ability to provide flexibility for diverse and evolving technologies,” Makowiecki says.

	In addition, a major trend in the cleanroom industry is a shift toward using modular construction, such as PODs, which are pre-assembled modules that are typically built offsite and then integrated into a facility onsite. PODs provide self-contained and autonomous cleanrooms (2).

	Reduction in lead time, increased flexibility, functionality, and ease-of-cleaning are factors driving the cleanroom industry toward modular systems (2).

	A critical point to consider when creating a modular manufacturing system is the biomanufacturing process that will be used with the system. To that end, GE Healthcare designed KUBio, a prefabricated, modular manufacturing facility solution based on single-use technologies, to optimize the process flow in biopharmaceutical production. The facility has been standardized to reduce facility-modelling time and offers a consistent setup in any global location.

	One of the challenges in the industry today is that the the large and diverse pipeline of biologics does not largely fit the current manufacturing infrastructure, Makowiecki points out. Higher product titers, biosimilars and new personalized therapies are driving batch sizes, cost reduction, and multiproduct manufacture flexibility. 

	“KUBio has been created with these drivers in mind, providing flexibility, reduced time to production, defined investment from the beginning of the project and global regulatory compliance,” Makowiecki adds.

	POD cleanroom units differ from traditional, fixed cleanrooms in that they offer ease of scalability, are mobile, and can be repurposed once a production process reaches the end of its lifecycle (3). PODs can be applied to many types of biopharmaceutical product manufacturing, including monoclonal antibodies, vaccines, recombinant proteins, personalized medicines, cell therapies, and gene therapies. In addition, the unit can be used for laboratory work (4).

	G-CON Manufacturing, a provider of autonomous cleanroom PODs, works with pharmaceutical and biotechnology companies to design POD cleanrooms that fit the specific applications required by these companies. 

	Following construction of the initial box structure, G-CON would then install internal wall systems and outfit the unit with ductwork, airhandling systems, automation and controls, doors, windows, flooring, and fire alarm systems, depending on the specified design (4).

	Once built and outfitted, the PODs are subjected to a factory acceptance test (FAT) to confirm that predetermined specifications are met. From there, PODs are wrapped and shipped to the onsite location, where they will be interconnected, according to G-CON. The process of constructing PODs differs from the traditional method of constructing cleanrooms because PODS require less crew member and less time to construct (4).

	Because modular systems can also be integrated into already existing facilities, it offers a less capitalintensive means to adopt modular manufacturing than having to build a new facility. As an example, GE Healthcare’s FlexFactory, a configurable, integrated, single-use biomanufacturing platform, is designed to fit a new or an existing manufacturing plant. 

	“The FlexFactory is a flexible, multiproduct biomanufacturing solution that can incorporate existing, qualified technologies as well as integrate new technologies that improve costs and process efficiencies,” says Makowiecki.

	The KUBio facility consists of 50–80 prefabricated modules, each module manufactured and fitted out with building services infrastructure, including heating, ventilation, and air conditioning (HVAC), electrical, and plumbing prior to shipping.

	“Parallel production pathways save time; while the modules are being manufactured, the customer site is prepared, and the manufacturing equipment is secured,” according to Makowiecki.

	Furthermore, a prefabricated facility like KUBio can be added to an existing site to increase manufacturing capacity or to increase production in a segregated manufacturing environment for sensitive products such as viral vectors that require an enhanced biosafety level of production.

	Adding capacity is particular challenging for pharmaceutical facility planners because it involves many strategic engineering and logistical factors. Modular manufacturing is an optimal platform to address these challenges by helping facility engineers maintain timelines and cost certainty while allowing them to adhere to design specifications (5).

	Modular pharmaceutical manufacturing helps mitigate the problems in older, traditional biomanufacturing buildings, many of which were designed prior to the most current cGMP regulations. These traditional facilities often have energy-inefficient heating, ventilation, and air conditioning (HVAC) systems that may not be properly isolated from other operations, according to Germfree Laboratories, a provider of custom-built biosafetly equipment.

	Modular manufacturing systems offer solutions to problems such as these by providing self-contained, energy-efficient HVAC and air filtration systems. They also give engineers a new range of options and allow engineers to focus on changing product requirements, capacity demands, and new R&D initiatives during the planning and design phase (5).

	“We see that there is an increasing need for flexible, multiproduct, prefabricated biomanufacturing solutions for commercial production. Today there are three KUBios and over 50 FlexFactories globally. Most of these facilities are used for clinical production, but there are also commercial sites, and the number is expected to increase in the coming years,” says Makowiecki.

	1. R. Hernandez, BioPharm International, 28 (5) 18–25 (2015).
	2. C. Lipeles and P. Genois, “

,” accessed 20 July, 2018.
	3. G-CON Manufacturing, “

,” accessed 20 July, 2018.
	4. S. Sarkar, “

Changing the Face of Biopharmaceutical Cleanroom Infrastructures

,” accessed 20 July 2018.
	5. Germfree Laboratories, “


	Vol. 30, No. 8
	August 2018
	Page: 30–31

	When referring to this article, please cite it as F. Mirasol, “Modular Manufacturing Can Ease Bioprocessing Woes," 

Modular manufacturing systems offer a less costly way to increase capacity while reducing time-to-market.

Modular Manufacturing Can Ease Bioprocessing Woes

As the pipeline of biologics matures, the pharmaceutical industry is expected to see new biotherapeutics that offer improved efficacy. Pressure from the current healthcare environment, however, is pushing the pharmaceutical industry toward lower-cost therapies and faster times to market.

	Biologic drug development has traditionally depended on the use of Chinese hamster ovary (CHO) cells. Over the past three decades, the CHO cell line has become the automatic choice of expression system for biomanufacturers, notes Mark Emalfarb, CEO of Dyadic International, a biotechnology company focused on developing biologic vaccines and biologic drugs.

	Emalfarb notes that the pharmaceutical industry is nonetheless experiencing the limitations that CHO expression yields present, which is complicated by pressures to develop affordable drugs quickly. “These [limitations] are particularly evident for the next wave of biologics, bi-specific, and tri-specific antibodies. The relatively low yields and relatively high costs of producing biologics with CHO appear to make it less sustainable and less commercially affordable for producing these more complex molecules,” Emalfarb says. “In a way, using CHO cells to develop and manufacture biologics is a bit like driving a 1900s model car in a 2020 Tesla world.”

	Using CHO cells as the industry standard for biologic drug development has several major drawbacks, adds Mario DiPaola, PhD, senior scientific director at Charles River Laboratories, a contract research organization. The most significant drawback is product yield, which is typically less than one gram per liter, followed by costs related to the extensive cell-line development and limited cell viability for production.

	“CHO expression systems work reasonably well for expression of less complex glycoproteins, but not well for new product modalities. For instance, in the case of AAV [adeno-associated virus]-like particle-based products, many developers are relying on HEK293 cell lines. Aside from yield and cost issues, the CHO cell-based expression system has other limitations related to glycosylation,” DiPaola explains. “The introduction of the immunogenic Galactose-alpha 1,3–Galactose linkage or N-glycolylneuraminic acid into glycan moieties of glycoproteins lacks certain transferases such as alpha 2,6 sialyltransferase, and it fails to produce any glycan with alpha 2,6 sialic acid.”

	Driving innovation behind better biologic drug development is decreasing the cost of these therapies, which is quite high for most biologic drugs, DiPaola comments. “It is important to drive the costs down through innovations in order to lower both the costs of development and costs of production. It is interesting to note that over the course of the past two decades the time from patent filing to market has increased from an average of 130 months to about 150 months, and the cost of getting a biologic approved has gone from approximately $650 in 1996 to more than $2 billion in 2018,” DiPaola says. “Clearly, these trends must be reversed so that more patients can benefit from these drugs at reasonable costs.”

	In addition, there is a separate need for more complex drugs, including more effective antibody drug conjugates (ADCs), multi-valent antibodies, and gene/cell therapies, “all of which industry to ensure success,” according to DiPaolo.

	Emalfarb further comments to say that “‘better, faster, more affordable’ is a mantra we should all take to heart as we pursue the knowledge that could-at least potentially-make it easier to make the drugs that could save lives more affordably.”

	“It is the responsibility of those working in the industry to enhance pharma companies’ ability to develop and make drugs efficiently and more cost-effectively. A large part of this entails encouraging pharmaceutical companies to study the potential of viable alternatives to CHO and to 

). Working together, government, regulatory agencies, academia, and pharma and biotech companies can indeed change things for the better,” Emalfarb says. 

	A cell source being explored as the basis of a new expression system for biologic building blocks is the fungus 

, nicknamed C1, which Dyadic has been developing for the past two decades. C1 technology is a fungal expression system for gene discovery, development, and production of enzymes and other proteins.

	Known as the C1 Expression System, Dyadic’s technology turns genes into a broad range of products and helps to overcome some of the inadequacies inherent in existing expression technologies used for gene discovery, product development, and commercialization.

	“C1 scales up the rate at which enzymes and other proteins can be produced and was initially used to produce biofuel and other enzymes in greater amounts and in less time than was possible before. To develop C1, our scientists exposed the cells of the 

species of fungus to ultraviolet light. They expanded and reinforced beneficial mutations to change the shape of C1 from long, spaghetti-like strands to short, grain-sized sections. Since C1 fungal cells secrete enzymes from the ends of the filaments, there were more secreting ends, multiplying the potential total yield of proteins,” says Emalfarb.

	“Furthermore, due to its new shape, C1 became easier to grow in large tanks. It offers a much shorter production time for monoclonal antibodies (mAbs) than CHO, requires smaller production facilities, and does not require expensive production media nor viral purification. When C1-expressed proteins are secreted from the cells, they come out in a purer form than CHO-produced mAbs,” he adds.

	For those reasons and others, C1 is said to represent an “innovation” in biologic drug development. “We hope one day C1 will be a safe and efficient expression system that can help speed up the development, lower the production costs, and improve the performance of biologic vaccines and drugs at flexible commercial scales. We believe it may also potentially enable the development and commercialization of genes that are difficult to express at reasonable yields in CHO, 

, and other cell lines and might also be able to produce larger amounts of protein to enable drug discovery and development to move forward faster,” Emalfarb states.

	Research results to date support the use of the C1 expression platform on an industrial scale in the biopharmaceutical industry. “In January 2018, for example, the maximum mAb yield from C1 stood at 1.34 grams per liter per day (G/L/d). Two months later, we had raised that to 1.71 G/L/d,” Emalfarb explained, then continued, “As of May 2018, this figure had been raised again to 2.46 G/L/d-representing an 84% improvement in productivity in only four months. This progress is accompanied by a corresponding 67% drop in the cost of the already low-cost, chemically defined medium required to produce the mAbs” (1,2).

	Charles River, which worked on approximately 80% of the drugs approved by FDA last year, has many technologies and methodologies that can be applied throughout the development cycle of a biologic drug, according to DiPaola. The company specifically develops a wide spectrum of animal models for testing products against specific diseases to screen, confirm, or validate a product’s functionality and biological activity in vivo in the early stages of development.

	“Charles River also possesses a variety of in vitro screening assays to assist in identifying the most potent drug candidates. To help drug developers weed out toxic drug candidates early in development, Charles River offers toxicological studies based on robust protocols utilizing multiple species,” DiPaolo says. The company also offers capabilities that support production cell-line characterization, cell banking, and viral clearance for purification-process validation.

	The C1 expression platform offers the biopharmaceutical industry potentially significant time and cost savings at each stage of cell development and biomanufacturing process development when compared to CHO cells, Emalfarb remarks.

	For one, C1 creates a stable cell line in a shortened timeline. “Dyadic has estimated a reduction of 50% in the time from gene fragment to stable cell line; C1 stable cell lines can be developed in just over three months versus six-plus months typically needed for CHO,” he says.

	Other “savings” benefits of the C1 expression system, according to Emalfarb, include:

		Significant upstream bioprocessing savings: C1’s doubling time (rate of cell growth is two hours versus 20 hours for CHO cells), leading to an estimated 10-day savings in time for the cell growth phase to charging the production fermenter with the required cell densities needed for commercial biologics production.

		Greater savings once commercial fermentation starts: The production time for C1 is significantly shorter-four to seven days versus CHO, which is typically a 12- to 14-day process, offering a 50% or even greater time savings.

		Saving on cell media costs: Based on the large difference of media needed for production using CHO, C1 achieves higher productivity using low-cost-defined synthetic media, estimated to offer a cost savings of five to 10 times over CHO media.

		Additional downstream processing savings: These are expected when using C1 versus CHO including the elimination of two viral inactivation steps that are typical in CHO biomanufacturing.					 	

	“As indicated earlier, C1 shaves off 50% or even greater time savings in the commercial fermenters, but most importantly, C1’s productivity is higher despite the much shorter time needed for production of biologics,” says Emalfarb.

	Dyadic recently reported that the company was able to produce a fulllength mAb with productivity of nine grams per liter in 90 hours, or in less than four days (1). This represents a greater-than-twice the industry average productivity for CHO, which is reported to be approximately 4 G/L, in one-third the time. “Putting this in perspective, C1’s mAb productivity equates to 2.4 G/L/d versus .30 G/L/d for CHO using much lower-cost media and the elimination of two viral purification steps typical for CHO,” Emalfarb notes.

	The most pressing challenges that the biopharmaceutical industry faces are development cost and time, both of which are becoming unsustainable for the industry, DiPaolo states. Another significant challenge is the lack of internal innovation and research capabilities across many of the medium-to-large size biopharmaceutical companies. “As a result, they must rely primarily on collaboration, in-licensing, or acquisition of new start-ups for innovation and new product leads,” DiPaolo says.

	“Some significant re-engineering is needed within the industry to bring about new strategies and approaches, possibly through greater use of automation, robotics, and artificial intelligence to allow for faster transition of biologic drug candidates from lab bench to clinic and then from clinic to market. These changes are necessary in order to address unmet medical needs, such as treatment of types of cancers, Alzheimer’s disease, and certain autoimmune diseases in a quicker and financially efficient manner,” DiPaola asserts.

	1. Dyadic, “Results Highlighted in Bioprocess International and the Global Bioprocessing and Bioanalytics Conference 2018,” Press Release, June 18, 2018.
	2. Dyadic, “Dyadic-C1 Technology,” presentation at Global Bioprocessing and Bioanalytics Conference (Prague, Czech Republic, 2018).


	Vol. 42, No. 8
	August 2018
	Page: 16–19

	When referring to this article, please cite it as F.Mirasol, “The Challenge of Building Better Biologic Drugs," 

Development costs and time to market continue to put pressure on the biopharma industry, driving the need for innovation in methods and technologies.

The Challenge of Building Better Biologic Drugs

From boosting cell-culture capability to growing bioreactor capacity, contract manufacturing organizations (CMOs) have been making investments to expand biologic output. 

highlights some of the major biologics-based CMO activity in the past months.

	In March 2018, Fujifilm acquired Irvine Scientific Sales Company and IS Japan, both cell-culture media companies, in a stock purchase agreement valued at around $800 million. The acquisition expanded Fujifilm’s biopharmaceuticals product portfolio to include in-vitro fertilization and cell therapy. The company stated that the acquisitions will expand its biopharmaceuticals contract development and manufacturing business and accelerate regenerative-medicine R&D as well as expand its reagent business. The company also plans to establish a new site in Boston, MA, by March 2019 for marketing products and services that support R&D and new-drug manufacturing (1).

	In November 2017, Celonic, a biologics development and manufacturing services provider, acquired a biomanufacturing facility in Heidelberg, Germany, from Glycotope, a clinical-stage immune-oncology company. With the acquisition, Celonic expands its GMP manufacturing assets, including continuous perfusion manufacturing, which the company now offers in addition to fed-batch manufacturing and the ability to scale-up from pilot to commercial-scale production. Combining Glycotope’s GlycoExpress human cell-line platform with Celonic’s CHOvolution integrated development platform will create custom-tailored expression systems (2).

	In October 2017, Lonza Pharm & Biotech acquired a clinical-stage mammalian manufacturing facility in Hayward, CA, from Shire. The 58,000-ft2 site includes 1000-L and 2000-L single-use bioreactors and associated downstream capabilities and provides added cGMP capacity to supplement existing assets in Lonza’s Slough, Berkshire, United Kingdom site (3).

	Lonza plans to add mid-scale mammalian capacity at its facility in Portsmouth, NH, with the installation of multiple 6000-L bioreactors, the company announced in May 2018. Construction is expected to start in late 2018. The company will also include the addition of multiple cell-therapy suites as part of the expansion. The hybrid facility will house full-suite process analytic technology, multi-variate analysis, and single-use technologies in an existing building and is expected to be operational by late 2018 (4).

	Patheon, now a part of Thermo Fisher Scientific, is expanding its biologic drug substance manufacturing capabilities with a $50-million investment by Thermo Fisher into the Patheon Biologics Center of Excellence (COE) for Biological Commercial Manufacturing in St. Louis, MO. The site provides process development, clinical cGMP manufacturing, and commercial manufacturing using fed batch and perfusion processes. The investment, announced in April 2018, includes a 64,000-ft2 expansion to the existing manufacturing building #2 at the site and will feature Thermo Fisher bioproduction bioreactors, consumables, and factory automation. The expansion will double the site’s manufacturing capacity. The expanded site is expected to be operational in 2019 (5).

	WuXi Biologics, part of WuXi AppTech, announced a couple of expansion plans in recent months. In April 2018, the company said it plans to invest €325 million (US$381 million) in a new biomanufacturing facility in Mullagharlin, Dundalk, County Louth, Ireland. This new facility will use multiple single-use bioreactors for commercial biomanufacturing and is designed to be able to run continuous bioprocessing. The company intends to install a total of 48,000 L fed-batch and 6,000 L perfusion bioreactor capacity (6).

	Prior to that, in March 2018, the company announced that it would install 4000-L custom single run (CSR) disposable bioreactors from ABEC, a provider of integrated process solutions and biopharmaceutical manufacturing services, at its new 30,000-L commercial manufacturing facility in Wuxi City, Jiangsu, China. The 4000-L CSRs will enable scale-up of manufacturing up to 24,000 L (7). The new 500,000-ft2 Wuxi City facility, in which the company invested $150 million, became fully operational in December 2017. It quintuples the company’s existing manufacturing capability and will support the biologics commercial manufacturing pipeline coming from global partners. Following completion of the first phase of construction in September 2016, the facility offers two 1000-Ldisposable bioreactors for perfusion processes, making it the largest perfusion biomanufacturing facility using disposable bioreactors in Asia, as reported by WuXi Biologics. The facility was later equipped with an additional 14 2000-L disposable bioreactors for fed-batch cell culture (8). 

	AGC Biologics, a contract development and manufacturing organization (CDMO), is adding a 2000-L single-use bioreactor to its Berkeley, CA, facility and plans to build a new complex that will house the company’s headquarters in Bothell, WA. The bioreactor addition at its Berkeley facility will support the company’s growing biologics capacity, which has tripled over the past three years. The Berkeley facility uses both single-use and stainless-steel bioreactors for cell-culture manufacturing in scales of 100–3000 L (9). The new 15,000-ft2 complex to be built at Bothell will provide expansion space for additional manufacturing capacity as well as in-house process development labs and corporate administrative offices. The Bothell expansion will also include a new R&D center dedicated to novel therapeutic protein manufacturing technologies (10). The company announced both these investments in March 2018.

	CMO Emergent BioSolutions is equipping its bulk manufacturing facility in Bayview, Baltimore, MD, with a 4000-L custom single run (CSR) bioreactor supplied by ABEC, a provider of engineering, equipment, and services to the biopharmaceutical manufacturing sector. The company announced the addition in October 2017. The 4000-L bioreactor, designed specifically for Emergent’s process needs, operational requirements, and facility constraints, will expand cell-culture capacity at the Bayview site and is expected to improve process transfer and scale-up, according to ABEC (11).

	1. Fujifilm, “Fujifilm to Acquire Irvine Scientific Sales Company and IS Japan, Leading Companies of Cell Culture Media,” Press Release, March 29, 2018.
	2. Celonic, “Celonic Acquires Glycotope’s Bio-manufacturing Facility,” Press Release, Nov. 3, 2017.
	3. Lonza, “Lonza to Acquire a US Clinical Manufacturing Site to Support Ongoing Customer Demand,” Press Release, Oct. 3, 2017.
	4. Lonza, “Lonza to Invest in Portsmouth, NH (USA) Site, Including Mid-Scale Biologics,” Press Release, May 4, 2018.
	5. Thermo Fisher Scientific, “Thermo Fisher Scientific Announces $50-Million Investment to Enhance Biologics Capabilities,” Press Release, April 23, 2018.
	6. WuXi Biologics, “WuXi Biologics to Invest €325-Million to Build Largest Biomanufacturing Facility Using Single-Use Bioreactors in Ireland,” Press Release, April 30, 2018.
	7. WuXi Biologics, “WuXi Biologics to Install Industry’s Largest Single-Use Bioreactor from ABEC at its New Commercial Manufacturing Facility,” Press Release, March 20, 2018.
	8. WuXi Biologics, “WuXi Biologics Initiated cGMP Manufacturing in the World’s Largest Biologics Manufacturing Facility Using Only Single-Use Bioreactors,” Press Release, Dec. 6, 2017.
	9. AGC Biologics, “AGC Biologics Expands Capacity at Berkeley, California Facility,” Press Release, March 6, 2018. 
	10. AGC Biologics, “AGC Biologics Increases Footprint in Bothell, WA,” Press Release, March 29, 2018.
	11. ABEC, “ABEC 4,000-Liter Single-Use Bioreactor to Expand Emergent BioSolutions Manufacturing Capability,” Press Release, Oct. 24, 2017.

	When referring to this article, please cite it as F.Mirasol, " CMO Roundup: Expansions for Biomanufacturing," 

Pharmaceutical Technology Outsourcing Resources Supplement 

CMOs have been active over the past year in expanding their biologics production and capabilities.