Susan Haigney is lead editor of Pharmaceutical Technology and Pharmaceutical Technology Europe.

Susan Haigney

spoke with Lisa Crandall, MS, associate director CMC Project Management, and Peter Angus, PhD, director of Pharmaceutical Sciences, at Velesco Pharma, which provides a full range of stability testing of API and drug products, about best practices in stability testing.

 Which methods are used to determine stability of a drug in early development? In later-stage development? Do these methods differ?

 In early development, typical API and drug product stability analysis include appearance, assay, related substances, and water. Additionally, depending upon type of drug product formulation being assessed, tests such as assay of preservatives and/or antioxidants, pH, viscosity, osmolality, particulates, antimicrobial effectiveness, and microbiological bioburden may be necessary. Specific methods such as high-performance liquid chromatography (HPLC) for assay/impurities are phase appropriate.  In later development, the stability indicating techniques haven’t changed, but the specific methods have evolved with the product.  Quantitation of known impurities is now possible.

What steps should be taken to ensure stability of a drug?

 To increase the chances that your drug is stable, it is vital to pay attention to all early data pertaining to the effects of temperature, humidity, light, polymorph, salt form, etc., as well as excipient compatibility and packaging. Discovering limitations and interactions early in development provides a better chance of success, especially in situations in which one polymorph is significantly more stable than others.

 What mistakes do companies make in early development that could lead to instability?

 Common mistakes early in development are ignoring the importance of risk assessment using the philosophies in International Council for Harmonization (ICH) Q8 and Q9, and not allowing enough time for designing a quality product. With appropriate foreknowledge, many drug instability challenges can be overcome by designing the synthetic route, formulation, packaging, storage, etc., with the future in mind.

What are some best practices for ensuring product stability in both early and later-stage development?

 To ensure the best chance for a successful and stable product, it is important to design early phase experiments to provide as much understanding about the API and product as possible. As the product progresses, it is important to use and build upon the data in hand to make necessary adjustments to synthetic routes, formulations, manufacturing processes, packaging, and analytical methods.

	When referring to this article, please cite it as S. Haigney, “Determining Drug Stability," 

Articles

Pharmaceutical Technology spoke with Lisa Crandall, MS, associate director CMC Project Management, and Peter Angus, PhD, director of Pharmaceutical Sciences, at Velesco Pharma, which provides a full range of stability testing of API and drug products, about best practices in stability testing.

Determining Drug Stability

Editor's Note: An abbreviated version of this article was published in 

Drug shortages are one of the most talked about topics in bio/pharma in recent years. External factors such as natural disasters and regulatory pressures, over which pharmaceutical companies might have little control, can contribute to drug shortages. Internal factors, such as failure in process controls, can and should be addressed by the pharmaceutical industry. Robust process development can help ensure that life-saving medicines reach patients.

	Chris Moreton of FinnBrit Consulting and Susan J. Schniepp, executive vice-president of Post-Approval Pharma and Distinguished Fellow at Regulatory Compliance Associates, discuss how poor process design can affect the supply chain and the pitfalls some pharma companies fall into.

 What are some common pitfalls pharmaceutical manufacturers face when developing manufacturing processes for the manufacture of small- or large-molecule drugs?

Schniepp (Regulatory Compliance Associates):

 I think one of the major pitfalls that affects the development of a robust manufacturing process is consideration regarding the equipment used, not only in the development of the process, but also in the transfer of the process to various manufacturing sites. I think this pitfall applies to both large- and small-molecule development. The new products are developed with consideration to the use of modern equipment. When these products get transferred from the development arena to the manufacturing arena, the equipment might not be comparable. The manufacturing facility may have older versions of the necessary equipment for producing the product, which could result in problems during transfer of the process. In some cases, the older equipment may require more human interaction, which could compromise the ability to meet the operating requirements specified in the original process. This would prompt changes to the original process that might significantly impact the efficiency of the manufacturing process and create deviations that need to be investigated to assure the product is safe to be released. Some process changes could also result in regulatory changes requiring prior approval before they can be implemented.

 I cannot comment on big pharma, but for small pharma/start-ups working with small molecules, common pitfalls include insufficient understanding of the limitations of the API, excipients, and processing. The questions that should be asked for any pharmaceutical formulation and process development project are:

		What do we have to do to get this drug substance into a form that will provide acceptable bioavailability via the intended route of administration?

 How can poor process development affect the supply chain?

Poor process development will most likely lead to stoppages, failed batches, and possibly reduced shelf-life. If the drug product cannot be made on a routine basis, there will likely be supply chain interruptions and patients will suffer. This can apply to both clinical supply and commercial manufacture.

 Poor processes generate deviations, which need to be investigated before product can be released. The more deviations there are associated with the product, the more chance the supply chain will be interrupted. If the original deviations are not investigated thoroughly, [there is the] chance that repeat deviations will occur. The more deviations associated with the process indicates deficiencies in the process. Until these deficiencies are successfully resolved, there is a good chance that the supply chain will be disrupted.

 How can poor process development affect a company’s bottom line?

 Poor process development leads to inefficiencies in manufacturing. These inefficiencies lead to delays in product release as well as product rejection. Unless the inefficiencies are effectively resolved and remediated, the company will continue to experience delays in product release and lost revenue from product rejection.

 Problems with supply of the drug product, for whatever reason, will mean that patients may to transferred to alternative products, possibly permanently. This will result in lost sales and thus lost revenue.

	During clinical trials, interruptions to the supply of an investigational drug can compromise study results, cause delays in completion of clinical trials, and possibly delay the filing of the marketing application, and thus loss of future revenue.

 How often should process development be reassessed? How often does a process change within the lifecycle of a drug product?

 There really is no hard and fast rule to how often process development information should be reassessed. Process deviations, manufacturing equipment changes and investigation results can prompt process changes. The process development information should be used to assure these changes are acceptable to make and do not compromise the quality of the product. Process development information should not stand alone but should be used to support the continuous manufacturing of the product over the product lifecycle.

 Typically, for small-molecule drugs, manufacturing processes for APIs may change, usually to improve yields or reduce the use of toxic solvents, etc. However, this is not so common because the impact on the final drug product performance must be assessed before the change can be implemented.

	In my experience, it is very unusual to make changes to a drug product manufacturing process. If they are made, the SUPAC guidance documents (Scale-Up and Post-Approval Changes) would apply in the United States (1–3). Similar criteria would be applied in the European Union.

	Looking forward, as quality by design becomes more prevalent, with the move to continuous verification, and with the current emphasis on quality metrics, more reasons for change may be identified as there will be increased scrutiny of data.

 Are there new trends in process development? Are companies moving to continuous manufacturing? 

 For small-molecule drugs, there is interest in continuous processing. However, it will not be for every product. Smaller-volume products will continue to be batch processed because of the changeover times. It can take up to a week to clean everything associated with a continuous processing line and be ready to run the next product. Thus, continuous processing is really better suited to larger volume products.

 I think the application of quality risk management as part of development activities is one of the major trends for process development. While the concept is not new, it is being applied more routinely in the process development phase to establish appropriate specifications, identify critical process parameters, and establish manufacturing controls. Using information from pharmaceutical development studies potentially helps identify control variations that may arise during processing.

Guidance for Industry: Immediate Release Solid Oral Dosage Forms Scale-Up and Postapproval Changes: Chemistry, Manufacturing, and Controls, 

Guidance for Industry: SUPAC-MR: Modified Release Solid Oral Dosage Forms Scale-Up and Postapproval Changes: Chemistry, Manufacturing, and Controls; 

Guidance for Industry: Nonsterile Semisolid Dosage Forms Scale-Up and Postapproval Changes: Chemistry, Manufacturing, and Controls; In Vitro Release Testing and 


	Volume 31, No. 1
	January 2019
	Page: 42

	When referring to this article, please cite it as S. Haigney, "Proper Process Development and Drug Shortages," 

Shortages of life-saving drugs are a regulatory and industry concern. Proper process development may help to ensure drug supply.

Proper Process Development and Drug Shortages

Contract manufacturing organizations (CMOs) and contract research and development organizations (CDMOs) invested in expanded facilities and services in late 2018. The following are some recent investments for biologic drug development and manufacturing.

	AGC Biologics, a clinical and commercial manufacturer of therapeutic proteins, announced on Sept. 20, 2018 that it will establish a new process development and manufacturing facility at its CDMO facility in Chiba, Japan, as part of its ongoing program to expand its production capacities globally (1). The new facility will contain single-use bioreactors at the 500- and 2000-L scale and will be suited for the production of monoclonal antibodies (mAbs), fusion proteins, and other types of therapeutic proteins. The facility is expected to be operational in the second half of 2019.

	Fujifilm Diosynth Biotechnologies (FDB) announced on Nov. 1, 2018 that its collaboration with UK-based Centre for Process Innovation (CPI), for the project AMECRYS: Revolutionizing Downstream Processing of Monoclonal Antibodies by Continuous Template-Assisted Membrane Crystallisation, has hit an important milestone with the successful technology transfer of the expression and purification of model mAbs from FDB to CPI (2). AMECRYS is a research project funded by the European Commission under the Horizon 2020 program in the framework of Future and Emerging Technologies actions (FET-OPEN), which supports early stages of science and technology research and innovation.

	Charles River Laboratories International announced on Oct. 25, 2018 that it has entered into an exclusive partnership with Distributed Bio, a company specializing in computational design and optimization of antibody discovery platforms, that grants Charles River access to Distributed Bio’s antibody libraries and integrated antibody optimization technologies (3). Distributed Bio’s libraries are computationally optimized for sequence diversity and engineering fitness through the analysis of thousands of human antibody repertoires and all known monoclonal therapeutics in clinical trials. The companies expect to create an end-to-end platform for therapeutic antibody discovery and development.

	ADC Biotechnology (ADC Bio) has secured additional funding of $3.18 million (£2.5 million) from existing investors and company management to ensure the achievement of specific business goals within the company’s overarching corporate strategy. 

	“We are delighted to have obtained this additional injection of funds that will be used to support the company’s strategic aspirations, including conceptual design of a downstream formulation and filling operation to complement our existing bioconjugation operations at the Deeside facility. We are also looking to fully exploit our core Lock-Release technology to create a transformative manufacturing paradigm that will significantly streamline ADC manufacturing supply chains,” said Charlie Johnson, CEO of ADC Bio, in a company press statement (4).

	1. AGC Biologics, “AGC Biologics Adds Mammalian Production Capacity in Japan,” 

www.agcbio.com/resource-center/news/agc-biologics-adds-mammalian-production-capacity-in-japan

 
	2. Fujifilm, “Fujifilm Announces that AMECRYS Collaboration Reaches Project Milestone to Develop New Technologies to Improve Downstream Processing of Monoclonal Antibodies,” Press Release, Nov. 1, 2018, 

www.fujifilmusa.com/press/news/display_news?newsID=881545


	3.Charles River, “Charles River and Distributed Bio Enter Exclusive Partnership to Create an Integrated Antibody Discover and Development Platform,” Press Release, Oct. 25, 2018, 

http://ir.criver.com/phoenix.zhtml?c=121668&p=irol-newsArticle&ID=2373437


	4. ADC Bio, “ADC Bio Secures Additional Equity Round Investment,” Press Release, Nov. 14, 2018, 


	Vol. 42, Issue 12
	December 2018
	Pages: 46–47

	When referring to this article, please cite it as S. Haigney, "Contract Organizations Expanded in Autumn," 

CMOs and CDMOs made investments in new and expanded facilities and services in the last quarter of 2018.

Contract Organizations Expanded in Autumn

	FDA Commissioner Scott Gottlieb has been promoting drug market competition in recent months that includes new guidance documents and targeted advisories to support R&D of complex drugs and combination products. A revised draft guidance discusses studies to evaluate the adhesive performance of proposed generic transdermal and topical delivery systems (TDS) products. A second guidance discusses studies to evaluate 

skin irritation and sensitization potential of such treatments. FDA also has issued 235 product-specific guidance documents on the appropriate methods for developing specific generic TDS products (1).

	In Europe, the European Medicines Agency (EMA) has pharmaceutical companies concerned after the agency announced it was slowing down work on certain guidelines due to Brexit. The industry has been waiting for a guideline on the assessment of device components in combination products under the European Union’s Medical Devices Regulation (MDR), which requires drug-device components in integral drug-device combinations (DDCs) to be evaluated by notified bodies. The process for this is still unclear, and some companies are worried this guideline delay will affect marketing authorization of integral DDCs, which range from pre-filled syringes to complex electro-mechanical devices (2).  

	EMA, however, has assured industry that work on the MDR guideline will continue. In an interview with 

, an agency spokesperson stated: “EMA is aware of the concerns raised by the industry that came with the introduction of the new MDR, and in particular with Article 117 (requiring an NB opinion on combination products) … Work on seven guidelines which address either an urgent public/animal health need or are necessary to support and facilitate preparation for Brexit or the implementation of new or revised legislation will continue beyond Nov. 1, 2018. This includes the guideline on quality requirements of medicinal products containing a device component for delivery or use of the medical product” (2).

	1. J. Wechsler, “FDA Promotes Complex Generics and Combination Products,” 

www.pharmtech.com/fda-promotes-complex-generics-and-combination-products-0


	Vol. 42, No. 12
	December 2018
	Pages: 39

	When referring to this article, please cite it as S. Haigney, “Regulatory Developments in Combination Products," 

FDA Commissioner Scott Gottlieb has been promoting drug market competition in recent months that includes new guidance documents and targeted advisories to support R&D of complex drugs and combination products. 

Regulatory Developments in Combination Products

 An abbreviated version of this article appeard in the October 2018 print issues of 

	An API’s physiochemical properties and its pharmacokinetic profile, as well as patient considerations, should dictate drug product formulation, according to experts at Catalent. Budget and timeline constraints, however, sometimes create difficulties. “The positive effects of formulation approaches on solubility, permeability, and ultimately bioavailability should be weighed against complexity, cost, and risk-to-launch of the chosen technology,” says experts at Catalent. “It is unrealistic to expect any formulation group to tackle all these considerations without the experience from many multiple product launches or the ability to leverage the expertise across a large and diverse formulation team.”

	According to Joe Masi, Sr. Director MS&T, at Cambrex, pharmaceutical companies turn to contract development and manufacturing organizations (CDMOs) to “resolve capacity shortages, tighten development timelines, reduce processing costs, and/or lack of internal development capability, etc.” Pharmaceutical companies are requesting end-to-end services more and more, according to Masi. “This includes API, formulation development, analytical methods development, manufacturing, and packaging development. In addition, pediatric formulation and fixed-dose combination products (two or more active ingredients in one product), as well as modified-and controlled-release complex formulation, continue to gain popularity and are often outsourced.”

	When it comes to outsourcing formulation development, however, challenges may arise when scaling up from small-scale batch to commercial production. Dr. Baerbel Hinneburg, director Technology and Process Transfer at Vetter Pharma-Fertigung GmbH & Co. KG, states that “concrete planning of execution with attention to detail is critical.”

spoke with Masi, Hinneburg, and experts at Catalent about the formulation and development issues that should be considered when addressing scale-up from small-scale batches to commercial production.

	Moving from clinical to commercial phases

What formulation challenges occur when moving from clinical to commercial phases?

 From a processing and technical point of view, one example is a change in material and equipment that may occur when moving from clinical to commercial manufacturing, such as the use of larger compounding equipment or a change from disposable to non-disposable material. One must be aware of the impact a change in material could have on the relevant attributes derived from the drug product profile. This awareness avoids further lab trials that need to be undertaken to determine the appropriate operational parameters that help maintain the quality and functionality of the drug product producedwith the new process.

 Usually, batch size and equipment used throughout development phases are small due to API availability, manufacturing cost, and the scale needed to meet clinical and registration requirements. However, some manufacturing process parameters may need to be changed when scaling up or using large-sized equipment for commercial production.

	Common challenges could occur during different manufacturing steps. A few examples are listed below.

	Blending step: Material flow (i.e., the flowability of granules) is one common challenge during manufacturing. Funnel flow is non-uniform, and the materials adhere to the walls of the hopper, resulting in blend uniformity issues during the blend. To overcome the issue, change the geometry of mixers, blenders, and hoppers to improve flow of materials through the hopper. Another way is using vibratory mechanisms to ensure a mass flow or having a paddle stirrer in the hopper.

	Compression step: Sticking and capping issues are commonly observed during compression. When different compression machines are used, they may not directly generate expected results. Modification of a tableting process can sometimes reduce or eliminate film formation or sticking during compression without making any drug formulation changes. Modifications include changes to pre-compression force, compression force, and tableting turret dwell time/speed. These modifications may be helpful in delaying the sticking behavior.

	Coating step: When using a large-sized coater, some of the parameters from the small coater may not work and coating uniformity may suffer. Coating variability usually increases at a faster pace with higher pan speeds. Therefore, the first consideration is to reduce coating pace to obtain better coating uniformity. The spray distribution across the tablet bed may be another cause of the coating uniformity issue. However, with functional coatings it is important that each nozzle is spraying the same amount of coating suspension. Each nozzle must have an even spray and be calibrated to ensure it functions properly. In early formulation and clinical phases of development, there are options to modify the qualitative formulation to overcome these challenges. However, because it is often difficult to make major changes at later phase without regulatory involvement, engage with an experienced CDMO from the earlier clinical development phase. They can help to develop and manufacture quality products with minimal to no clinical or regulatory impact.

 The main challenge of a formulation proven as safe and effective for the therapeutic action tested in patients is to ensure that as we move from the beginning of the quality-by-design (QbD) process to commercial process validation, there are no changes in correlated critical material attributes (CMAs) (APIs, excipients, synthesis route, suppliers, etc.) and that none of the critical process parameters (CPPs) (associated with scale up to commercial batches) will affect the critical quality attributes (CQA) of the product that ensured efficacy and safety in clinical-phase stages. If any change is necessary to apply as part of the process, a risk assessment and mitigation should be implemented to assure the desired quality, considering the safety and efficacy of the pharmaceutical form.

	Critical quality attributes and critical process parameters

What steps should be taken for successful scale-up from small-scale batches to commercial production?

 To prepare a robust and reproducible commercial production process, we perform a QbD approach. This approach involves a combination of gap and risk analysis to identify and evaluate any factors that could potentially impact CQA and any not obvious scale-up process steps that become CPPs. A comprehensive process design to accommodate both known and newly identified CPPs, combined with a process qualification to verify a constant product quality and define a control strategy, is essential.

 First, define the target product profile (TPP), which describes the use, safety, and efficacy of the product. Prior knowledge and in-depth understanding of formulation, excipients, and process is advantageous when defining the TPP and will reduce the number of experiments and analytical testing required and, consequently, the manufacturing and testing costs.

	The next step is to identify the CQAs of the final product. CQAs should be studied thoroughly and controlled to meet the TPP. To achieve the desirable CQAs, it is necessary to identify and control CPPs. CPPs identified throughout the development and scale-up process include raw material and API controls (particle size distribution, polymorphs, and impurities), process controls, and design spaces around individual or multiple unit operations (granulation, compression, coating, packaging). These CPPs are monitored throughout development and updated upon the collection of new information.

	Successful scale up can be achieved by a QbD approach, which includes design of experiments (DoE), risk assessment, and process analytical technology (PAT).

 Most frequently, there will be changes between the equipment used in small-scale batches to ones in the commercial setting. With the difference in the equipment, the CQAs (e.g., dissolution) of the drug product could be affected, and this may depend on the complexity of the formulation. It is important to understand the correlations between the equipment scale, CPPs, and CQAs as this knowledge will help to fine tune the CPPs in the commercial-scale production that will produce drug product with the desired CQAs. These relationships can be studied by appropriate DoE at small-scale.

 How can quality by design be used to ensure quality during formulation development?

 The concept of QbD is to deliver a quality pharmaceutical product by designing a robust manufacturing process.

	Quality cannot be tested into the product at the end; it should be built into the product from the beginning. The knowledge gained from development studies can be utilized to design the working space, specifications, and manufacturing controls. Changes in formulation and manufacturing processes during development can be used to gain knowledge and support design space.

	Pharmaceutical QbD is a systematic approach with predefined objectives and emphasizes product- and process-understanding and control based on scientific knowledge and quality risk management.

	Successful QbD involves defining the target product profile and then identifying critical quality attributes, followed by developing and controlling critical process parameters etc., which will eventually ensure quality products.

	Solid QbD during formulation development can: reduce batch-to-batch variability and defects, improve product development and manufacturing efficiency, develop a robust manufacturing process that leads to greater regulatory confidence and increased product quality, improve yields, reduce investigations and testing, and lower manufacturing costs.

 Practicing QbD in formulation development emphasizes holistic and systematic investigation, understanding, and prediction of variances caused by the drug substance(s), functional excipients, formulation design, biopharmaceutics, and process development (instead of finding them out through normal means or through accidental variation). The key elements of QbD are risk assessment, conducted by cross functional teams; DoE; statistical analysis; and PAT. These elements help identify and measure risks and variabilities, correlate CMAs and CPPs to CQAs, establishing design space, control strategies, and relevant specifications, and ensuring right-first-time production at reduced costs, reduced time, and increased efficiency.

 How do analytical methods change from clinical development to commercial production?

 Often, analytic methods do not change dramatically from clinical development to commercial production, but the understanding of the method and information changes. Early phase methods are developed for speed and to minimize cost, and the assay/purity methods are commonly adapted from the method developed by the API manufacturer (for ease of tracking API impurities).

	Dissolution methods may simply be discriminating rather than profile generating, and things such as extraction procedures may need to be optimized. Once a drug has moved from clinical to commercial, a commercial quality control laboratory performs a method evaluation and transfer to confirm that the method can be performed, and the same results can be obtained using similar instrumentation. If there are any nuances to the method (understanding its variability, the level of validation performed, the experience on different equipment or by different analysts), then changes are usually minor adjustments and the method is updated. Changes in the analytical methods could support certain adjustments during a validated-state maintenance process if they merit it. Historical data generated by the method can be used to adjust or refine the acceptance criteria as the program progresses from clinical to commercial batch production.

	It is key to use these data to assess critical method parameters that must be controlled carefully as part of the overall analytical control strategy-as the molecule moves to commercial testing and release.

	Statistical tools are valuable to set acceptance appropriately for commercial products-while also considering practicality, so as to not fail a batch unnecessarily.

When moving from clinical-scale production to commercial production, what validation steps must be performed?

 The successful transfer of a product from clinical- to commercial-scale production is based on a thorough understanding of the manufacturing process, the inherent variability in the process, and strategies to mitigate or control these sources of variability.

	This knowledge is gained through scientifically based process development work and documented in reports that are used as the source documentation to create the commercial validation plan.

	The validation plan and process risk assessments are used to justify and implement the validation strategy, number of validations batches to be executed, sampling plans, and testing criteria.

	The validation batches are executed under protocol by trained personnel using qualified equipment. Enhanced physical and analytical testing may be done to assure process robustness and control. A validation summary report including physical and analytical batch data, statistical data treatment, and summary of batch outcomes is approved by discipline subject matter experts and the quality unit prior to commercial batch release to distribution.

 A total of three consecutive, successful (commercial-scale) batches need to be manufactured within 10 times the size of the registered batch size. Validation demonstrates that a specific process will produce batches that meet specification and that normal variation would not predict an out-of-specification result. Emphasis is given to those elements that have been established, through QbD, as having a significant impact upon product quality, accompanied by increased testing of samples from throughout the process. It is not good practice to use validation batches for experimentation beyond that which has already been demonstrated, as the costs of validation batches are typically very high.

What are some best practices for successful tech transfer?

 In our experience, a dedicated transfer team that includes a wide breadth of experts is crucial. This team is responsible for the process design required to perform a QbD-driven tech transfer. Roles and responsibilities must be agreed upon, and a system that enables adequate communication and feedback should be established. Open communication and exchange of all information gained during development is a key element. The license holder should also check early in the process that all partners and suppliers can provide adequate quality and documentation systems that help ascertain regulatory requirements are being met.

 The main goal of tech transfer is to transfer the product and process with minimal or no changes, which will minimize regulatory challenges and smooth the path to regulatory approval.

	The success of a technology transfer depends on several things: the quality of the finished product, open communication between two parties, feasibility of scale-up to desired levels, and compatibility of equipment at the transferred site. Therefore, it is advisable to consult with the technical and regulatory experts from the transferred site regarding the feasibility of the process with minimal impact on finished product.

	Important actions to take for a successful tech transfer include:

	• Obtain detailed technology transfer documents such as product development reports, batch records, protocols, and documents containing CPPs, CQAs, and TPPs from the transferring site. Better communication between transferring and transferred site is a key for successful tech transfer.

	• Understand formulation, manufacturing process, key equipment, function of each and every excipient, specifications, and critical manufacturing process parameters etc. for the tech transfer product.

	• Perform a gap analysis between sites (transferring and transferred site) by evaluating the equipment and supporting the information by comparing differences in the make, model, type, and capabilities of equipment available between transferring and transferred site.

	• Identify the regulatory strategy; SUPAC guidelines describe equipment in detail and classifies changes in three levels: Level I, Level II, and Level III changes. CBE30, PAS, and annual reportable are common strategies for tech transfer, which can save companies significant time and money.

	• Perform feasibility batches and capture the critical process parameters and optimize the process before registration/validation batches.

	• Gather stability data including bulk hold data on finished product to gain more confidence on the quality of the product from transferred site.

	• Generate a comparison report to compare equipment and manufacturing process parameters between transferring and transferred site and to perform a risk assessment.

 First, understand and capture the historical technical details or lessons learned from previous manufacturer (s) via discussions or detailed development reports.

	Second, understand customers’ timelines for milestones and plan critical activities (e.g., raw materials, specifications, analytical method transfer/validation, and ancillary equipment parts) accordingly.


	Vol. 42, No. 10
	October 2018
	Pages: 68–70

	When referring to this article, please cite it as S. Haigney, “Outsourcing Development: Small-Scale to Commercial," 

Industry experts discuss the formulation and development issues that should be considered when addressing scale up from small-scale batches to commercial production.

Outsourcing Development: Small-Scale to Commercial

 spoke with Aled Jones, senior product and applications manager at ProZyme; Philip Widdowson, European Application Development Scientist at Thermo Fisher Scientific; and Daryl Fernandes, chief executive at Ludger Ltd about the challenges of performing glycan analysis and how outsourcing companies can offer specific expertise.

What are the specific challenges associated with glycan analysis?

Thorough drug glycosylation analysis would involve full structural characterization of the oligosaccharides of each unique glycoform as well their absolute quantitation. However, this is not straightforward and getting even close to this still requires significant time, resources, and skill. To achieve this ideal you would, in principle, need to isolate each glycoform (including nonglycosylated drug variants), determine its relative abundance, then elucidate the glycan structures at each glycosylation site. However, it’s generally very difficult to separate out all of a drug’s glycoforms with complete resolution, and even if you have isolated a single glycoform, you need to deal with the glycans.

	At first glance, glycans look like they should be simple to analyze because they’re small molecules with a relatively small number of constituent monomers.

	However, unlike polymerization of monomers in linear macromolecules, there are many ways to link monosaccharides together to form glycan moieties. So, for each glycosidic linkage, you need to determine the identities of the connected monosaccharides, the anomericity (i.e., the special configuration) of that bond, and which of the hydroxyl groups around the monosaccharide rings are involved in the linkage.

	Also, most drug glycans are not linear structures but are branched. This means you can find glycans with identical masses and monosaccharide constituents but different topologies, and these topological isomers can confer different physicochemical and biological properties to the drug. Another problem is skewing of glycoform profiles. This can be caused by degradation of glycans or either selective losses or enrichment of glycan species during sample preparation.

	A further issue is that underivatized glycans can be difficult to visualize at analytical scale. They don’t have natural fluorophores or strong chromophores for HPCL [high-performance liquid chromatography], and their ionization on MS [mass spectrometry] can be challenging as the glycan signals are readily attenuated by other chemicals such as salts and peptides.

As glycan analysis is still somewhat of a niche, it’s perhaps less accessible than genomics or proteomics.

	Glycans display a broad structural diversity based on monosaccharide composition, linkage type, and branching. Added to this complexity is the site of glycan attachment; there is usually just one Nglycan site in the case of most IgG-based therapeutics but there can be more in Fc fusion proteins and other glycosylated biotherapeutics, and O-glycans may also be present in these.

Due to the nature of their production, glycoproteins

	are highly heterogeneous molecules with numerous different glycan structures present at a single glycosylation site (1). Glycans are commonly released from the protein prior to analysis; however, the chemical properties of glycans make them difficult to analyze in their native form.

	Therefore, glycans are required to be derivatized in order to make them amendable for traditional analytical methodologies (2). O-glycans are much more difficult to analyze in the released form due to the lack of a universal release enzyme (3). In addition, glycans can be isomeric in composition, which complicates the accurate determination of structure, particularly when using methods such as mass spectrometry (4).

What are the various methods for glycan analysis?

Glycan analysis can be approached on a few different levels, from intact protein through to glycopeptides, released glycans, right down to individual monosaccharide components. Each of these techniques have their advantages and drawbacks. The route taken may depend on the molecule and the data required, but in general, relative quantitation of released glycans labelled with a fluorophore and separated by liquid chromatography is one of the most common approaches for analysis of mAb [monoclonal antibody] Nglycosylation.

Depending on the specific information required, glycosylation analysis can be performed on different levels. Intact glycoprotein analysis allows for a quick overview of glycan distribution to be carried out. Analysis at the glycopeptide level makes site-specific glycosylation profiling possible by assigning specific glycan structures to distinct glycosylation sites.

	Analysis of released glycans analysis is currently the preferred strategy due to the high amount of information regarding the total glycan population, which can be elucidated (2). For released and fluorescently labeled glycans, HILIC [hydrophilic interaction liquid] chromatography is commonly applied either in isolation for relative quantification through fluorescent detection or coupled to mass spectrometry for characterization. More recently, there is a drive in the direction of analysis at the glycopeptide level, specifically when assessed as part of a multi-attribute method (MAM) workflow (5).

The key is to develop a glycoanalysis toolset composed of orthogonal techniques that reliably measure the glycosylation critical quality attributes (GCQAs) for your therapeutic. You need several methods because there isn’t a single technique that can cover all glycan analysis needs. In particular, you must have reliable methods for characterizing and quantifying potentially immunogenic glycans (such as those bearing Gal-alpha–1,3Gal motifs), as well as those for glycans that confer high therapeutic activity for your drug. So, selection of glycan analysis methods must be driven by a thorough understanding of the glycan structure-protein activity relationships for your therapeutic.

	Also, your toolbox must furnish the breadth, structural detail, and sensitivity needed for every stage of your drug’s lifecycle-and at suitable cost and throughput.

	This allows you to tune the methods according to the task. For example, glycan analysis for selection of clones to match your quality target product profile (QTPP) or in QbD [quality-by-design] studies to determine design space (DS) will typically require a very different performance profile from those needed for batch release.

What are the benefits of outsourcing glycan analysis for sponsor companies?

There is no single method that permits complete glycan analysis, and therefore, a panel of methodologies is often required not only to fully characterize the glycans, but also to meet regulatory requirements. Meeting these requirements in full demands the use of numerous types of instrumentation and analytical platforms. Sponsor companies regularly do not have direct access to all of these platforms, and it is not often economically or logistically viable to bring them on-site. Dedicated outsourcing companies for glycan analysis, therefore, play a key role in filling any potential gaps which may exist with respect to instrumentation and necessary expertise required to perform these methods (6).

Smaller or early-stage companies may not have the personnel to perform glycan analysis, or the instrumentation required such as liquid chromatography with fluorescence detection, capillary electrophoresis, and mass spectrometry.

	The existing instrumentation they have may not provide the data quality or capabilities of the ‘latest and greatest’.

	Or there simply may not be time to implement or run an analytical method for glycans within the constraints of a project timeline. In these cases, it can make sense for sponsors to seek an outsourcing partner that has experience with glycan analysis and can turn data around in a timely manner.

Outsourcing glycan analysis can allow sponsors to significantly extend their capabilities for drug realization. However, for success, it needs to be well-planned with a carefully chosen outsourcing partner. When poorly done, outsourcing can result in you losing time, money, and control of a critical part of your drug’s development and give you data that’s less than useful. Good outsourcing with an accomplished partner does the opposite, allowing you to overcome regulatory hurdles more smoothly and get your product to market faster than if you relied solely on your own resources.

	The first step to successful outsourcing is choosing a reliable partner with the expertise and services that match your needs.

	They need to demonstrate their ability and experience with therapeutics of similar or greater complexity to your drug. Other key considerations include the quality of communication between you and your outsourcing partner, whether or not they will share their detailed methods with you and what follow-on services they could provide. Not that reliable glycan analysis will typically be much more complex and therefore more costly than, for example, routine peptide analyses. However, with a suitable outsourcing partner and carefully considered and well-executed plan, you should benefit from a significant return on your investment.

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Considering the challenging nature of performing glycan analysis, what are the benefits of outsourcing glycan analysis for sponsor companies?