Cynthia A. Challener is a contributing editor to Pharmaceutical Technology.

Cynthia A. Challener

Conventional microbial analyses require extensive manual operations. In addition, the necessity for visual determination of the growth of microbial colonies leads to longer assay times. The result is a lengthy process that can take anywhere from three days to more than two weeks and that can dramatically impact production and commercialization times. Automated systems providing rapid results are helping to reduce that time, but the key to acceptance is staying consistent with the compendial method. Cynthia Challener, PhD, editor of the 

Pharmaceutical Sciences, Manufacturing & Marketplace Report

, spoke with Julie Sperry, chief commercial officer of Rapid Micro Biosystems, to gain further insight into the application of rapid microbiological testing.

Pharmaceutical Sciences, Manufacturing & Marketplace Report:

 For what applications in the pharmaceutical industry is rapid microbiological testing appropriate and why? What benefits/advantages does it provide? 

There are many areas where microbial analysis is critical in pharmaceutical manufacturing. Raw materials, work-in-process material, and final products all must undergo microbial testing. In addition, microbial monitoring of the manufacturing environment (i.e., personnel, surfaces, and air) is an ongoing process and critical to compliance. As a result, pharmaceutical quality-control laboratories often process hundreds, if not thousands, of tests per day, much of which involves manual manipulation of the assay plates for reading as well as data logging and other activities.

Rapid analysis with automation can, therefore, reduce the burden of manual plate counting and data entry on these laboratories and allow the workforce to be redeployed to address other important tasks. In addition, the level of human error can be reduced, and because the automation can provide the data immediately, decisions can be made quickly and definitive actions on process development issues and product releases can occur sooner.

Elements of a rapid microbiological system

 What are the critical factors that contribute to a successful rapid microbiological analysis system? 

 The most critical factor is the use of the established, or compendial, method, so that the analytical method doesn’t involve a major process change and can be validated without any need for extensive validation work.

Second, it is important that the method is nondestructive. If contamination is detected, then the microbes must be identified. Therefore, the test must be designed so that it doesn’t destroy or cause any change in the microbes. There are several ‘alternative’ technologies available, but many involve destruction of the microbes. In addition, many of these other methods don’t report the results in colony-forming units (CFUs), which is the standard unit for microbial testing. Therefore, the data have to be related to CFUs, which require validation of the data correlation and can introduce uncertainty.

: How is it possible to develop an automated system that can meet these requirements?

The key is to use a growth-based method but [also to] develop a visual analysis system that can detect colony formation much sooner than is possible with the human eye. Our system is based on the natural auto-fluorescence of the microbes. Samples are prepared using media that is typically used for conventional analyses. A certain wavelength of light is shone on the sample, and if there is microbial contamination, the microbes auto-fluoresce. A very sensitive camera detects and captures this fluorescence.

To establish the growth rules for the software, extensive tests were conducted on many different types of microbes, including bacteria, molds and yeast, in order to develop reliable software that can detect colony growth and disregard non-growing debris, such as lysed cells or particulate components. The system was validated by comparing the results of automated determinations with determinations made using the conventional method.

The system takes readings every four hours and looks for differences. Often contamination can be seen within 8-12 hours, so in many cases, a problem can be identified after just two to three readings. The system then sends an alert message indicating that the colonies detected have exceeded a preset alert limit. With this type of automated detection system, it is possible to reduce the time needed for microbial analysis to about half of what is required for the conventional method.

 How is automated microbiological testing being improved? What recent advances have been made?

 One of the issues for some of our pharmaceutical customers is the need for separate systems for sterile-product testing. Others, on the other hand, want a system that can handle the whole range of different analyses, including sterility, bioburden, and environmental monitoring.

We have taken these customer requests very seriously and have developed a line of second-generation products, which will be released mid-year, that address these issues. One of the new instruments is specifically for high-throughput environmental monitoring and water/ bioburden analyses and is designed for high throughput.

The second new system is a specialized instrument for use in sterility-testing applications. This system includes the ability to test for both aerobic and anaerobic microbes. The system has three cassettes, one of which creates an anaerobic environment with a system sensor to detect and confirm that the cassette maintains an anaerobic atmosphere throughout the testing. The emphasis here is not only on time to results, but also proper sample preparation and handling.

The third new system is a multifunctional instrument designed to handle all of the different types of microbial analyses required in pharmaceutical manufacturing and whch can simultaneously run multiple analyses: sterility, environmental monitoring, water, or bioburden. This system is useful for smaller companies that can’t afford several individual instruments, but is also suitable for larger manufacturers that want to have the flexibility to validate a range of applications and then spin the validation out to their various sites.

Most importantly, the same growth-based method and detection technology is used across the product line. One added feature, though, is that these systems can be connected to a laboratory information-management system (LIMS) for improved sample tracking.

 What limitations remain with respect to rapid microbiological testing? What might be done to address these challenges? 

 There are several issues related to sample preparation and the subsequent identification testing that play a role in the turnaround time for the overall microbial testing process. Logistics, for example, can account for a significant amount of time and resource consumption. Often the testing is done in one location, but samples are collected elsewhere. Pharmaceutical companies can make efficiency gains with automated systems that can be operated in or closer to the manufacturing facility, particularly for environmental monitoring analyses. The capabilities of our new system facilitate this testing shift. Longer term, we are interested in finding ways to help our pharmaceutical customers speed up sample preparation and microbe identification so that they can maximize the increases in productivity and decision-making that rapid, automated microbial analysis systems can provide.

Articles

It can take up to two weeks or more to obtain results from traditional microbiological testing methods. Rapid, automated testing can reduce that time in half.

Advances in Rapid Microbiological Testing

A high proportion (> 50%) of the active pharmaceutical ingredients (APIs) approved by FDA during the past 10 years were manufactured using chiral technology, according to industry estimates. Asymmetric transformations are thus a critical aspect of drug manufacturing today, and catalysis has become the primary method for the efficient and selective synthesis of single enantiomer products. Development chemists now have a choice between enzyme-based biocatalysts and transition metal-based chemocatalysts (some organocatalysts are attracting interest as well but are not discussed in this article).

 Chris Senanayake, vice-president of chemical development in the US for Boehringer Ingelheim Pharmaceuticals, Ian C. Lennon, senior vice-president for global business development with Chiral Quest, Antonio Zanotti-Gerosa, team leader with Johnson Matthey Catalysis and Chiral Technologies, Ephraim Honig, chief operating officer with Strem Chemicals, and Wataru Kuriyama, a chemist with Takasago International, discuss important aspects of chemocatalysis for asymmetric synthesis.

Pharmaceutical Sciences, Manufacturing and Marketplace Report: 

Considering the state-of-the-art catalytic toolbox of today, when are homogeneous chemocatalysts advantageous over other types of catalysts for chiral transformations?

In preparing the initial batches of API for preclinical studies and early clinical trials, the simplest, not necessarily the best and most economical, method is used due to time constraints. That may be a well-known stoichiometric method using a chiral auxiliary or a resolution-based approach. In parallel, we investigate chemocatalytic routes to determine if there are any cost-effective, scalable alternatives, considering all aspects, such as the metal (availability, cost), ligand (intellectual-property issues, cost), yields, selectivities, and catalyst loadings. Enzymatic routes generally take much longer to develop and often require the use of outside expertise for the engineering of the desired reactivity, so they tend to be used more often in second-generation processes in late-stage drug development.

 Chemocatalysts can be classified into two major types, homogeneous catalysts and heterogeneous catalysts. Heterogeneous catalysts have advantages in recovery and reusability; however, generally, homogeneous catalysts show better reactivity and selectivity. In addition, the flexibility of ligand design can improve selectivity, and in asymmetric reactions, high selectivity is required. Therefore, homogeneous catalysts are predominantly used. 

Zanotti-Gerosa (Johnson Matthey Catalysis and Chiral Technologies):

 The use of biocatalysis is growing, but the choice of catalyst technology must be made on a case-by-case basis. Chemical catalysts offer some real advantages. Many are available off-the-shelf and do not need to be engineered for a specific reaction. They have high activity, often require only a simple workup, and most importantly, have high-volume efficiency. What we see is that the speed of development that is possible with chemocatalysis is also very attractive because it enables more rapid development of production solutions. All of these factors have to be considered in order to find the most cost-effective catalytic solution.

Choice of method—such as between chemocatalysis and biocatalysis—will also depend on the technology that is possessed by the CMO. Chiral Quest, for example, specializes in asymmetric hydrogenation.  There are also areas where biocatalysis is not yet effective, such as the asymmetric synthesis of α-amino acids. Asymmetric hydrogenation, on the other hand, is exceptionally good for the commercial-scale manufacture of both aliphatic and aromatic α-amino acids. 

Ligand, substrate, recovery and cost challenges 

Pharmaceutical Sciences, Manufacturing and Marketplace Report:

 What are the major shortcomings/limitations of current homogeneous chemocatalysts for chiral transformations? What major issues would you like to see addressed in next-generation catalyst technologies?

 The synthesis of chiral ligands is often very difficult and can require the use of cryogenic conditions and resolution steps.  On the positive side, in many cases, both enantiomers can be used, and the catalyst loadings are very low, so large quantities of the ligands are not needed. Obviously, having much cheaper and easier to make ligands while still maintaining excellent enantiomeric excess and high activity is a good goal.

On the substrate side, there are also still a number of compound classes that are challenging. For asymmetric hydrogenation, imines, oximes, 

-substituted double bonds, and aromatic systems fall into this category. Some advances are being made in the asymmetric hydrogenation of heterocycles, and this area holds much promise as it will lead to the manufacture of complex chiral molecules in very few steps and is a reaction that cannot be achieved using enzymes.

 Removal of the metal and reuse are issues that must be addressed, but there are processes where homogeneous catalysts are recycled. Rh-BINAP in our 

-menthol process, for example, is recycled from the distillation residue. However, there still is much room to improve metal and catalyst removal. Next-generation catalysts should have the advantage of both homogeneous and heterogeneous catalysis as well as high selectivity and recyclability.

 The catalyst cost is the biggest issue. Most chiral catalysts for asymmetric synthesis contain precious metals and very complex ligands, both of which are very expensive. Therefore, a cost-effective process requires very low catalyst loadings (ideally 100,000:1). Going to such low levels also eliminates the need to recover the catalyst.

Scalability is the biggest issue. Often with catalytic reactions, the catalyst does not seem to behave the same way on a very large scale as it does on the bench scale, and this issue can be more pronounced with asymmetric catalysis. This change in behavior can be due to catalyst stability issues or problems with the purity of the starting materials. Boehringer Ingelheim has established an in-house Catalysis Group to design new ligands and to develop new catalytic reactions that can address both the cost and scalability issues encountered in API development.

 What is being done to overcome these problems?

Zanotti-Gerosa (Johnson Matthey Catalysis and Chiral Technologies): 

Existing catalysts are being applied to other transformations, or being modified so that they can be effective for related reactions. For example, some hydrogenation catalysts have been shown to be effective for the reverse dehyrogenation reaction.  Other catalysts have been introduced that can do both hydrogenation and transfer hydrogenation reactions.  This flexibility has a cost impact. Development work is also proceeding with many important chiral catalysts and ligands whose patents have recently or will soon expire.

Immobilization is one solution for avoiding recovery issues. In general, immobilization can be achieved through binding of the ligands or the metal to a solid support. From the perspective of industry, immobilization of the metal is advantageous because ligands developed for homogeneous catalysts can be used as is, which reduces the cost and time of catalyst development. Rhodium (Rh) with the well-known Duphos ligand on PTA (phosphotungstic acid) for asymmetric hydrogenation is one example of this type of catalyst. Microencapsulation is another important method. Toxic OsO

 immobilized by this method is used for Sharpless asymmetric dihydroxylation and recovered without any leaching.

 In our experience, it is better to optimize the homogeneous reaction to get the lowest possible catalyst loading and the best selectivity, so that each production run takes the same time, gives the same enantiomeric excess (ee), uses the same catalyst loading, and provides the same purity of product. Consistency is especially important for GMP manufacture. Generally, making the ligand heterogeneous leads to poorer catalyst loadings (often 10 x higher or more), lower and less reproducible ee’s, and sometimes incomplete reactions.

 The key to developing a catalytic reaction into commercial scale is to gain an understanding of the mechanism of the reaction and identify the active catalytic species. It is imperative that the interactions between the catalyst and the substrate be identified. The use of high-throughput screening, combinatorial ligand discovery, computational tools, and modern analytical techniques in an integrated manner will make it possible to obtain the necessary level of understanding of catalytic behavior needed for the development of robust systems that will be effective at any scale.

 Of recently introduced chiral chemocatalysts and/or ligands, what do you believe are the most novel and why? Which ones will have the most impact on fine chemical/pharmaceutical synthesis and why? 

 We believe that our asymmetric hydrogenation catalysts RUCY and DENEB are the most novel catalysts and will have the most impact on fine chemicals. RUCY and DENEB show unprecedentedly high catalyst activity; the productivity per hour or per weight of catalyst is high enough that the cost for production can be reduced and the removal of the metal is relatively easy. In addition, RUCY and DENEB have a wide substrate scope and reduce substrates that can’t be hydrogenated by conventional catalysts. Therefore, we believe that they will have the most impact on the pharmaceutical industry, where efficient chemical transformation of compounds with a variety of functional groups is required and product quality must be tightly controlled.

 There is a lot of interest in iridium complexes developed by the Kerr group at the University of Strathclyde for the direct, flexible, and selective introduction of hydrogen isotopes to organic molecular frameworks under extremely mild reaction conditions, and these catalysts have successfully been used for the isotopic labeling of several known drug molecules. We also believe that the RUCY-XylBINAP catalyst developed by Takasago, which is effective at catalyst loadings as low as 0.001 mol%, is very promising as an industrially practical catalyst for asymmetric hydrogenation.

The Noyori ketone hydrogen technology, for which the Nobel Prize was awarded in 2001, is a very important asymmetric transformation today. In fact, second-generation catalysts that are more robust (more resistant to deactivation) and more active that are being developed by Walter Baratta at the University of Udine and others will be valuable new catalysts for pharmaceutical manufacturing. New catalysts for asymmetric transfer hydrogenation of ketones and imines being developed by Martin Wills at Warwick University are also noteworthy. 

 Asymmetric hydrogenations are widely used in industrial pharmaceutical synthesis and are least affected by scale up issues. Jacobsen epoxidation chemistry, the Buchwald–Hartwig aminations, Suzuki and other cross-coupling reactions as well as transfer-hydrogenation processes are also important transformations that are being developed on larger scales.

The ligands that will have the greatest impact on fine chemical/pharmaceutical synthesis are those that are being used today. For example, Chiral Quest manufactures many tons of products using our asymmetric hydrogenation catalysts, such as Rh-DuanPhos and Ru-C3-TunePhos. There are so many useful ligands and catalysts available that any new system will find it hard to compete.  A new ligand would need to be very easy to make, very active, and highly enantioselective or carry out a reaction that none of the existing technology can achieve.

Although the use of biocatalysis for asymmetric transformations is growing, chemocatalysis remains a highly competitive technology.

Asymmetric Chemocatalysis: Going for the Lowest Loadings

Continuous manufacturing has been common practice in many industries for decades, but it is just now entering the early phases of adoption in the pharmaceutical industry. With effective online analytical capabilities available, equipment suppliers have been able to develop systems for various aspects of the tableting process, and drug manufacturers are beginning to realize the many benefits of continuous processes. Although challenges remain, expectations are high for the eventual fuller use of integrated continuous processes for solid-dosage manufacturing.


There are several benefits associated with effective continuous production systems. Increased safety and lower costs are on top of the list. “Continuous processes are more integrated with fewer steps, which leads to less manual handling and increased safety,” says Kris Schoeters, product manager continuous processing for GEA Pharma Systems. “In addition, the equipment and facilities are much smaller, allowing flexible operation at lower capital costs, with less work-in-progress materials.” Along with the monitoring and tracking, which are a fundamental aspect of continuous manufacturing, there also is better control of the process than is possible with batch processes, so both the consistency and quality of tablets are improved, according to Stephen Boswell, director of S3 Process, a provider of process equipment.  

Additionally, the development time and costs for continuous processes are often less than those for conventional batch-manufacturing operations. “Inherent R&D advantages include quicker capabilities to test a variety of process conditions and the use of smaller quantities of expensive API,” notes Sharon Nowak, global business development manager for food and pharmaceuticals with K-Tron Pitman. Adds Ivo Backx, manager of business and project development for the pharmaceutical industry with Siemens: “The time is shrunk at the development stage and during the move from development to production, in part, because process analytical technology (PAT) tools are an integral part of the development process and also because there is no need for scale-up to different equipment. Commercial-scale quantities can be produced on the same continuous tableting equipment used during the development stage.”

Benefits continue to accrue after product launch as well. Capacity can be dynamically adjusted to meet market requirements by running a process for longer or shorter periods, according to Backx. Boswell adds that with smaller equipment and less capital investment, it is also possible to locate continuous tableting lines in a greater number of geographic locations so that transportation costs are lower and drugs can be formulated for the local market. “Such advantages are particularly critical for US and European manufacturers to remain competitive,” he notes.

Overall, companies that implement continuous manufacturing processes expect a fast return on investment and cost savings from 10-20% compared to batch operations, according to Backx. Furthermore, with the reduced waste generation, greater product recovery, lower energy consumption, and overall improved operation efficiency, continuous processes enable “greener” manufacturing.


Even with these benefits, pharmaceutical companies have been unwilling until recently to make a move toward continuous tableting. “There has been a hesitancy to make investments in continuous manufacturing. It is quite a big step for drug manufacturers to take,” says Boswell. Some of that hesitancy, according to Schoeters, has been related to the need to establish that each batch of product, ultimately released following sampling and testing of finished product, is homogeneous. “Traditionally, this characteristic is established by blending the batch immediately prior to compression, but in a continuous process, the batch is never assembled in one place. Previously, the available on-line monitoring tools were not efficient enough.” 

Recent developments in the use of PAT for on-line and at-line monitoring to prove the quality of product and process continuously as well as the design and engineering of continuous process components in pharmaceutical execution, have helped pave the way for continuous processing consideration, according to Nowak.  

The introduction of equipment for flexible, compact, quasi-gram-scale continuous manufacturing has also been critical, according to Schoeters. GEA focused on integrating the entire process cycle from weighing, blending, granulation, and drying up to tablet compression in its systems. “Also important is a steady-state controllable flow of ingredients into the granulator, even in the face of often-challenging powder flow properties, and to ensure plug flow so that tracing of ingredients for specific ‘batches’ can be established.” Adds Boswell: “A substantial derisking of R&D scale-up is possible with certain continuous technologies, such as that from Gerteis, where process parameters are the same between clinical-trial to large-scale manufacturing, [thereby], saving substantial scale-up cost and risk.”

K-Tron also has addressed the need for equipment that can process smaller quantities of material for both development and pilot-plant continuous processes and for poorly flowing APIs and excipients. It has systems designed to deliver as little as 20 grams per hour with high accuracy, according to Nowak, and its latest feeder, which will be introduced at Interphex in April, functions effectively with difficult cohesive powders.

FDA’s support of continuous processing is also causing many pharmaceutical companies to seriously pursue this option in development and manufacture. Still, Backx warns that companies need to have a clear understanding of their business goals when implementing continuous tableting or any other continuing manufacturing operation. “For some companies, continuous manufacturing might provide a new approach to improving a problematic process through increased process understanding and control while others may be looking to first implement continuous manufacturing for drug-development efforts rather than changing existing processes.”


Of course, there are challenges still to be faced before continuous tableting will be widely adopted in the pharmaceutical industry. Nowak points to two major issues that are currently being addressed: the integration of the complete continuous process and defining both the actual lot size being produced and the size and frequency of sampling measurements.  

Schoeters agrees that full integration of all of the different PAT measurements is critical, as is intelligent feed-forward and feed-back loops and control systems to ensure the most efficient performance. 

K-Tron, which has experience with these types of online feed-loop controls in equipment for the plastics and food industries, expects that they can easily be modified for utilization in a continuous blending or granulation process to assist in the overall quality of the product and process, according to Nowak.  “The ability to monitor material feed properties and integrate this data into a sophisticated overall data management tool will not only provide for enhanced process understanding, but also enable end-users to specifically understand the process impacts of different lots of raw materials.” 

There are also implications for facility design that must be considered. “The workflow within facilities must change with the adoption of complete continuous tableting processes,” says Boswell. “Those changes won’t happen overnight, but they will happen over time.”

Regulatory-related issues are causing some companies to remain cautious. The uncertainty regarding how regulators will formalize the approval of continuous manufacturing and real-time release strategies is a concern, notes Backx. He points out, though, that the uncertainty is largely due to the lack of experience rather than any actual regulatory impediments. Some are also worried about continued emphasis on the term “batch” in regulations. “This concern is unfounded, however; regulatory definitions for the term ‘batch’ already include those batches produced through continuous methods—the mode of manufacturing is not mentioned. Companies establish the ‘specific quantity’ of output that will count as one batch for tracking and tracing and potential recall purposes,” Backx explains.


As pharmaceutical companies begin to realize the significant potential for cost savings and quality improvements, many of these issues and concerns will be addressed. Companies, such as Siemens, GEA Pharma Systems, K-Tron, and S3 Process, are working closely with customers in the pharmaceutical industry, software platform manufacturers, and universities to gain greater process understanding and develop fully integrated continuous tableting systems that will ultimately include data-driven quality verification for in-line assurance and real-time product release.  

Equipment suppliers are helping the pharmaceutical industry move towards adoption of continuous tablet production.

Making Strides in Continuous Tableting

Biopharmaceutical manufacturers are looking to their raw-material suppliers to provide them with the support of a secure supply chain. To gain a perspective, Cynthia Challener, PhD and editor of the 

Pharmaceutical Sciences, Manufacturing, and Marketplace Report

, recently spoke to Beth Ann McLane, SAFC’s product manager for enhanced quality, of the importance of ensuring supply-chain transparency and quality for raw materials. SAFC recently introduced a line of PharmaGrade products for raw materials for biopharmaceutical applications. 

Public demand for increased supply chain security

What situation led to the decision to develop your new PharmaGrade raw materials for the biopharmaceutical industry?

 Biopharmaceutical manufacturers are faced, now more than ever, with the reality of global events that have threatened and breached supply-chain security. These occurrences have caused a public outcry, and regulatory bodies have demanded higher levels of compliance. Manufacturers are now being held fully responsible for the raw materials that go into their products, and they simply do not have enough capacity to test for all of the possible contaminants that could find their way into the materials. 

In response, the industry is moving towards risk assessment and risk mitigation of raw materials from the early stages; in essence, manufacturers are looking to their raw-material suppliers to provide them with the support of a secure supply chain. The more that biopharmaceutical manufacturers know about their suppliers and the raw-material supply chain, the more confident they can then be about the safety of those raw materials.

 How is the introduction of a new grade of pharmaceutical raw materials anticipated to fill these market needs?

 PharmaGrade products were developed to provide better supply-chain transparency and manufacturing controls; they are a lower-risk option to use in manufacturing than many currently available raw materials. The portfolio was created in response to urgent demand from customers for products they can count on to be consistent and that are from a source where the production is controlled and changes are limited. These types of customers are not only looking for a guaranteed level of quality, but they also want to know what is happening with the raw materials and to be notified if/when any changes occur.

When we started to build the product line, we anticipated that customers would be looking for a higher level of documentation and support than ever before. In response to this expectation, we have put a great amount of resources into identifying and qualifying products for the line. In addition, SAFC has a strong change control notification system and offers robust support, especially for our GMP and Elite quality-level products.The Elite products provide an option for customers that are looking for highly controlled products, but do not necessarily need them to meet the same level of validation as required by GMP guidelines.

Ensuring audit-worthiness of key ingredients

 What are the specifications/requirements for the new grade of raw materials? How were these determined? How are they different from existing raw materials?

 SAFC’s goal is to hit seven key criteria for the new materials that were directly based off of customer requests. The first six are:

Manufactured under GMP (or SAFC’s Elite) quality guidelines

Quality assurance review and release of each lot

Readily available quality documentation (laid out in an easy-to-use format)

The challenge for raw-material suppliers has been to produce and sell raw materials that can stand up to an audit. As such, one of our key goals is to make sure customers know what they are buying and are satisfied that the products meet their expectations for quality and documentation when they audit. The seventh, and perhaps, the most important criterion, is to have these products available as catalog items in inventory, so customers can have access to them when they need them.

Which types of materials is SAFC looking to offer at this new grade? How/why were they selected? 

 SAFC is offering a broad range of amino acids, buffers, and specialty chemicals as PharmaGrade materials. These chemicals were selected because they are components with known usage in biopharmaceutical manufacturing. We knew they would appeal to a broad number of customers.An example of a product we offer is L-methionine sulfoximine, which is used as a selection agent. Until SAFC introduced the PharmaGrade version of the product, the other products available in the market were not made at the same high quality level, under controlled processes, and with the same supply-chain transparency. Additionally, we wanted to be sure to offer commonly used chemicals, such as sodium butyrate, L-tyrosine, and disodium salt. 

 How has the introduction of the new grade of raw materials affected SAFC’s suppliers and customers? What feedback have you received?

 Many of our customers have been eagerly looking forward to the introduction of new PharmaGrade products, and the feedback has been very positive. They have been extremely pleased with the readily available inventory of each of our products as well as the enhanced quality product documentation that comes with them.

Each product has a complete dossier of information about it, including the site of manufacture and quality systems associated with the product. We are directly responding to customer requests for products that are available when they need them and in the format they need them. Customers have clearly expressed that they want us to expand the offering as quickly as possible.

The only real issue we have had is delays in adding new products to the line due to requirements we expect from suppliers. This line of products requires that our suppliers provide us the detailed information about raw materials and procured products so that we can offer the full level of supply-chain transparency expected with the PharmaGrade portfolio. If we realize that a product cannot meet the standards, then we will not move forward in adding it to the line. Or if we realize that we need longer to verify the standards, then we will take the time to make sure that happens.

 What can be expected in the short-term and long-term with respect to the PharmaGrade products for biopharmaceutical manufacturing?

 SAFC offers PharmaGrade products globally, and we will continue to launch new products globally each quarter. Our immediate plan is to double the size of the PharmaGrade portfolio by the end of 2013. Our long-term goal is to create a full line of products for biopharmaceutical manufacturing with the right supply-chain transparency and control to help customers reduce the risk in their raw materials and, ultimately, their final products.

Ensuring the quality of raw materials is a crucial component of achieving product quality of APIs and finished drug products.

Strategies for Achieving Biopharmaceutical Supply-Chain Security

As the demand for larger amounts of stem cells and other fragile cells grows, biopharmaceutical companies are struggling with how to effectively produce these specialized biological materials in larger quantities. ATMI LifeSciences has developed a new reactor to meet that need. Product Director Matthieu Egloff spoke with the 

 about the special conditions required for the larger-scale production of fragile cells and how its new Xpansion reactor was designed to provide them.

A podcast version of the interview also may be found 

 Why is there is a need for special reactors that are suitable for the growth of cells and particularly for fragile cells. Also, what are the trends in biologic API manufacturing that have led your company to develop a special reactor system?

 First, I will start by dividing or categorizing the cell-therapy market. There is a lot of noise today in the cell-therapy and emerging field, and it is a promising new therapeutic area. In these markets, you have different kinds of applications, but you have also different kinds of sources for cells and so have different products based on these cells.  

From all of these kinds of cells, one that is today making a lot of noise is stem cells. You have different sources of stem cells, adherent stem cells and pluripotent stem cells. All of these stem cells are very fragile cells to grow and use, and the technology to cultivate these cells doesn’t really exist today, or at least in the last year, there were not many technologies able to grow these cells.

Because cell therapy is an emerging market, I think the main companies developing the biotechnology didn’t have the time to really get involved into this market and to develop the right technology. But, today, this dawn is changing, and biotech companies, such as ATMI, are now developing this specific bioreactor.

And I will say why we have to develop a new bioreactor. Before, most of the biotech production was based on cell lines, mammalian cell lines, for adherent cells, for example, and these cells were used for vaccine production or for protein production. In this case, you just need to grow the cells and you will use what will be secreted by the cells. There is not really a need to harvest the cells at the end of the process.

But with cell therapy, it’s completely different. Now, you will have to harvest the cells because the cells are the product. You have to grow the cells and then make sure to be able to get them. That’s the tricky point from a technology perspective, and from a biology perspective, these cells, the stem cells, for example, are very fragile cells, meaning that they are very sensitive to what we call the niche microenvironment. You really have to control this niche microenvironment to be sure that the stem cells that you will grow will behave as you want.

So that’s the reason that you have to develop a very specific bioreactor that will support the right growth for these cells.

 What exactly makes the cells fragile, and what are the special requirements that are needed to be able to work with those cells?

 The main reason that these cells are fragile is because they are very sensitive to what I mentioned before, to the niche microenvironment, meaning they are very sensitive to the physiochemical parameters of pH and dissolved oxygen, and they are very sensitive to shear stress. Also, different factors and parameters can, in fact, affect the differentiation of the cells. That’s something very specific to stem cells. The stem cells can differentiate into different lineages, so according to different factors, they can differentiate into one type of cell or another one. And that’s why it’s very complicated to cultivate these cells because you really need to [not only] grow them, but to drive them on the right path. You have to avoid differentiation, or at least avoid the differentiation you don’t want to have. That’s why you really need a system that will be able to control these niche microenvironments.

 What are some of the special requirements that allow you to do that then in a reactor system?

  The first one, if we are talking mainly about adherent stem cells, is to control the environment of the cells, so you will have to control the pH and you will have to control the dissolved oxygen level into your bioreactor. You will have also to control and to minimize the shear stress, meaning that you can’t use a bioreactor with high mixing and shear stress. You will have to protect the cells; that’s the main requirement you need.

And, of course, for the stem cells growing in adherence, you will have to provide a surface to help the cells to adhere to something, to the surface or the scaffold needed to preserve the stem cells and not leave the cells to differentiate into another type of cell you don’t want to have.

 Can you describe your Xpansion Bioreactor system and how you’ve incorporated these requirements? How it is different from a conventional reactor as far as the design and operation, and how does it fit in with the rest of the manufacturing operation?

 Today, most of the cell-therapy companies, or the companies developing products based on stem cells and adjuvant stem cells, are using tissue-culture flasks or multi-tray vessels to grow these cells. And these companies, because they are in the early R&D phase, or in clinical and early clinical trials, are using multi-tray stacks. These technologies have been used in the market for a very long time. They are robust, and these technologies—the tissue culture flask and the 2D and multi-tray stacks—are what we can call planar technology, meaning a 2D surface, a plate surface, and plastic. A cell-therapy company has to scale up the process, but it’s just impossible to have a commercialization process based on a tissue-culture flask or multi-tray stack technology because with this system, for the controls, you have to operate the system with manual operation and you have to operate the system under very aseptic conditions. There is a lot of risk to continue to use these technologies for high-volume manufacturing at the commercialization stage. Most of these cell-therapy companies have to develop and to scale up these small scale processes. For them, the main requirement, of course, is to be able to preserve the cell’s integrity, identity, and potency.

Because the stem cells are very fragile, and because these companies have developed the cells onto a 2D surface, a plastic 2D surface, they have to keep the same kind of microenvironment. Again, still for the same purpose, to preserve the sensitivity.That’s why, at ATMI, we developed something very similar to what people are used to using. We developed a multi-plate bioreactor, and that is unique on the market today. There is no other multi-plate bioreactor. We have really compacted the design of a multi-tray stack to keep the same philosophy. We continue to use a 2D surface and with plastic. We are using the same polystyrene material as in multi-tray stacks or tissue-culture flasks, and we are using the same kind of surface treatment. Thanks to that, we are really providing to the cells the same kind of environment as in a tissue-culture flask or in a multi-tray stack.

That is the first innovation we brought—to really compare the design.  In a typical multi-tray stack, you have a lot of space between two plates, and with the design of Xpansion, we really succeeded in avoiding the space between two plates. Turning to a multi-tray stack, you have a space between two plates of about 16 mm; with Xpansion, we have only 1.6 mm between two plates. We also succeeded in reducing the thickness of the plate; it is about 3 mm in the multi-tray stack, and with Xpansion, we have only 1 mm of thickness for the plate. Thanks to that, we really compacted the design, and with one Xpansion bioreactor, that’s the name of the bioreactor, Xpansion, with the Xpansion bioreactor with 200 plates in only one device, it is equivalent to 20 multi-tray stacks.

To give you an idea, a 10 multi-tray stack is about 4 meters high, and the Xpansion with 200 plates is below 1 meter high. It has really decreased the footprint, but it’s also decreased the number of operations because you don’t have to manipulate 20 devices; you only have to manipulate one device.

That was the first innovation—to keep the same kind of material, to preserve the niche microenvironment, and to compare the design.  The second point was to add all the advantages of a bioreactor, meaning the capacity to control pH, to control the dissolved oxygen, inside the bioreactor. We also have another feature, the capacity to observe the cells inside the bioreactor. We have a specific microscope that helps us to observe the cell density and the cell confluency inside the bioreactor.

The main value of this bioreactor is to help our customers to switch easily from multi-tray stacks or tissue-culture flasks into a bioreactor system. Thanks to that, they are able to scale up the process. Then, of course, they are able to improve the safety of the process because it’s a closed system. They also are able to decrease the cost of goods because you minimize the number of operations. You need less operators to operate the system, and they are able also to decrease the investment cost. Because, thanks to the system with the bioreactor, you will minimize the number of incubators that are required, and you will minimize the number of laminar flow hoods that are required with the previous technologies.

Because the bioreactor can be operated in a closed system, you will be able to work in a Class C environment, so you will definitely minimize the size of your facility, and the class form environment of your facility. Again, you will decrease the investment cost.

 Are you working on further developments for fragile-cell manufacturing? Are you looking to improve the reactor or introduce any other technologies that would be of benefit?

  Yes, definitely, because we think that Xpansion is a great technology to help our customers to move from the R&D process to clinical production, and then to start to have early commercialization or mixed-scale commercialization of production and sales. But, some companies will have to produce a huge number of cells per cell batch, and for this kind of process, we are evaluating what can be the next generation of bioreactors for stem-cell production of very, very large numbers of cells.

For that, there are two main options today that we are evaluating. The first one is microcarriers, and it’s very difficult to grow stem cells on microcarriers for different reasons, but one of them is mainly the shear stress on the cells and how to guarantee seeding of the microcarrier and so on, and how to scale up a microcarrier-based process.

For that, we have a specific bioreactor at ATMI that has a specific and very low shear stress, thanks to a very high mixing efficiency.  It’s a PadReactor, and it’s based on a patented mixing technology. That’s one method we are exploring—the use of microcarriers and this bioreactor. The second option we are evaluating is to use what we call a 3D scaffold that provides a lot of surface for the cells into a small volume, and thanks to the 3D technology, we may be able to produce a very high number of cells in a very small bioreactor. Those are the kinds of technologies that we are today evaluating for the future.

 Do you have any estimation on a time frame for when these technologies would be available?

 Today, we have the PadReactor technology available. We have already a lot of data using this technology and microcarriers for viral production. We are working with the top vaccine producers in the world with these technologies, so this technology is used a lot. We now have to adapt the microcarriers for fragile cells, and we are now starting to assemble some partnerships to explore how to grow stem cells with the PadReactor technology. So, hopefully, in the next year, we will have good data to show to our customers regarding this technology. And for the 3D technology, it really depends on how we will push forward the development; the ongoing development is quite confidential today, so it is difficult to give you an answer on the second option.

Matthieu Egloff, product director with ATMI LifeSciences, discusses the need for larger, special reactors that can provide the necessary conditions for the production of fragile cells on a larger scale.

New Reactor Technology Enables Larger Scale Manufacture of Fragile Cells


Catalysis is critical to the synthesis of pharmaceutical intermediates and APIs. Catalysts make important synthetic reactions proceed more rapidly and often enable the formation of complex structures in fewer steps with greater selectivity. Biocatalysts, typically enzymes, offer additional advantages. They catalyze reactions under mild conditions and thus consume less energy. They do not require the use of heavy or toxic metals, and in some cases enable the avoidance of hazardous raw materials. In addition, such reactions can often be carried out in water rather than organic solvents, and they tend to generate less waste.

Until recently, pharmaceutical companies tended to consider biocatalytic routes only in later development stages. “Biocatalysts have not frequently been considered in the design of manufacturing processes for drug candidates in early stage development, such as candidate selection in preclinical development, according to Daniel Mink, corporate scientist, biocatalysis, from DSM. “Generally, biocatalysis has been treated as a viable cost-saving alternative only in late-phase clinical stages (i.e., Phase II and III).

Biocatalysis is now being applied throughout the pharmaceutical development process, beginning with the synthesis of compounds for structure activity relationship studies through initial route development and for commercial scale production, according to Tom Moody, head of biocatalysis with the Almac Group. 


Recognizing the growing need for biocatalytic expertise, Almac established a biocatalysis group in 2007 that has since doubled in size. The company recently invested £2.5 million ($3.75 million) in biocatalyst R&D and launched its own selectAZyme biocatalyst technology platform. “The biocatalysis group is now an integral part of the Almac Sciences’ business unit, which focuses on the development phase. Furthermore, Almac has successfully introduced biocatalysis in manufacturing processes for candidates in preclinical development that immediately resulted in substantial savings in terms of processing time, quantity of starting materials required, and quality of end product, with no impact on the development time, according to Moody.

DSM, meanwhile, has a proven track record with over 30 commercial bioprocesses run on a multiton scale with experience in the manufacturing of intermediates, APIs, and enzymes, according to Mink. “As a result, this collaboration on biocatalysis fits strategically and technically for both companies. Our manufacturing assets are complementary, and together we can offer a true one-stop-shop for biocatalytic solutions.” 

 As importantly for Moody, with the partnership in place, Almac now has the opportunity to screen the collection of DSM enzymes at Almac, while DSM has access to Almac’s proprietary enzymes. “Together, we now offer the pharma industry the largest enzyme platform for structural diversification and commercial scale production,” he explains. Each company has the entire platform of enzymes in house and available for screening but will continue to handle the sale of enzymes or transformations individually. “With such a large platform that offers many complementary enzyme chemistries, we have a much higher chance of finding a hit as the result of our enzyme screens,” Moody states.

Specific enzyme technologies of note include pig liver esterase (PharmaPLE) for sterically demanding esters, aldolases for carbon–carbon bond formation, and epimerases and oxidoreductases that make it possible to investigate different chiral spaces, which is critically important in pharmaceutical synthesis.


The goal of this partnership is to increase the services that both companies can offer to their customers. “Via this agreement, we maintain our commitment to the future of pharma manufacturing, adding to our green-chemistry toolbox. Working with the Almac Group marks further advancement of DSM's stated strategy towards strategic partnerships to excel in providing new customer solutions,” says Alexander Wessels, president and CEO of DSM Pharmaceutical Products.Notably, the agreement is nonexclusive. “The relationship with the customer is very open, and it is the customer who drives the entire process,” Moody says. Almac will always discuss manufacturing options in good faith with the client. If the process fits within Almac’s scale of manufacture, Almac will likely keep the project. If it exceeds Almac’s capacity, Almac will and recommend DSM where appropriate. 

In addition, customers can choose from several different business models. “From the earliest stage of discussions, the clients’ strategy for commercial supply will be discussed and defined, and a range of options will be offered to them,” notes Mink. 

If a customer is only interested in material delivery, Almac or DSM, as appropriate, will develop the process and supply the product at an agreed price. Alternatively, customers can pay for the process development work and own the process intellectual property, but still have Almac or DSM manufacture the product. Clients can also choose to have full control over the process, own the enzyme for the particular transformation (such as one that has been evolved at the behest of the customer to perform specific chemistry using the customer’s substrate), and transfer the manufacture to an external third party. Customers that choose to scale up their processes with DSM will benefit from a seamless transition. “Almac has an experienced and dedicated technical transfer team that has already transferred multiton scale manufacturing processes to commercial facilities in Europe and the Far East,” Moody observes. “Furthermore,” he adds, “Almac and DSM share the same approach to process development and have complementary production capacities, which should make the transition even smoother.”


The collaboration, announced in the fall of 2012, has already born fruit. The first project, for which Almac developed the process and has proven it at the pilot scale, will shortly be transferred to DSM for commercial-scale production. While the companies cannot provide details of the API due to confidentiality agreements, Moody has indicated that the compound is a natural product, and the process involves a multistep synthesis.

“We are excited about this collaboration and the fact that it will help increase our chances of finding the right enzymes for our customers and make it possible to access new and varied structures,” says Moody. “Both Almac and DSM are and will remain very active in the discovery and development of new enzymes that will further widen the spectrum of bioreactions possible and offer even more diversity to the pharma industry, adds Mink.”

Collaboration provides pharmaceutical industry customers with access to enzyme expertise and assets resulting in cost-effective, green processing at all phases of development, from preclinical to commercial manufacturing scale.