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BioPharm International, June 2011 Issue (PDF)

Leadership, partnerships, and breakthroughs will converge at the Biotechnology Industry Organization's (BIO) International Convention this month in Washington, DC. Leaders from 65 countries across the globe, representing all fields of biotechnology, will come together to share their insights and forge new partnerships.  

It is significant that the convention will be in Washington, DC, this year because the time has come to take our advocacy in the nation's capital to a higher and more aggressive level. Biotechnology is crucial for developing new medical therapies, but at the same time, the industry is facing increasing uncertainty in rewards, thus reducing its appeal for sustained investments. The public and policymakers need to better understand and appreciate these challenges. 

Our industry is facing struggles that cannot be modified by any single approach. Instead, we must find ways to reduce regulatory risk and uncertainly, enhance reward, and reinforce the position of the industry as a major contributor to resolving society's most pressing needs. 

Thankfully, policymakers who recognize the value of biotechnology also recognize that the right policy environment can help the industry thrive. The ability of members of BIO to heal, fuel, and feed the world requires an economic and public-policy environment that supports their work. For instance, a balanced pathway to biosimilars will give biotech investors greater confidence in taking the financial risks of funding the research of biotech startups. 

Other BIO policy priorities include ensuring that the National Institutes of Health has sufficient funding to sustain the public-private collaboration that is transforming biomedical discoveries into innovative treatments for patients. Congress must also provide FDA with the resources it needs to keep pace with rapidly evolving biomedical science and fulfill its vital health and safety mission. Strengthening FDA's review process is a focus of our ongoing discussions with policymakers about the next renewal of the Prescription Drug User-Fee Act. Working closely with our members, BIO also will remain engaged as the US Department of Health and Human Services moves to implement the Patient Protection and Affordable Care Act. 

The Therapeutic Discovery Project (TDP) program, enacted in 2010, is an example of the type of policies necessary to spur continued medical innovation, while at the same time protecting and growing high-paying US jobs. The TDP program provided $1 billion in research grants and credits for small biotech companies pursuing new therapies for diseases, such as Alzheimer's, HIV/AIDs, Parkinson's, and Multiple Sclerosis. BIO is calling on Congress and the Obama administration to work together to extend and expand TDP to support continued American innovation and further accelerate the development of life-saving cures. 

Meanwhile, along with our state affiliates, we will continue to work with state governments seeking to grow their biotech sectors as part of their economic development and job-creation strategies. BIO looks forward to engaging with leaders in Washington to ensure that policies support an economic environment that encourages innovation. BIO will continue to champion the policies that enable our members to saves lives and transform our world. 

is president and CEO of the Biotechnology Industry Organization (BIO).

Guest Editorial

BIO's president and CEO provides insight leading up to the 2011 convention.

Policy and Innovation

The Chinese hamster ovary cell line (CHO cell) is considered by many as the workhorse of the biotechnology industry. It is a preferred cell for production of an ever-expanding array of engineered products that have therapeutic, diagnostic or other applications. The cell line not only provides a well characterized laboratory for performing genetic manipulations, but also offers an excellent expression host for production of engineered products. The glycosylation pattern of this mammalian cell is an advantage not offered by potentially more productive microbial expression systems. Inherent in its mammalian nature, however, is its relatively slow growth rate and the constraints on its cultivation.  

Although great improvements have been made in the culture of most animal cells, resulting in increases in their productivity, their use in large scale production has been limited in the past to batch or fed-batch processes. In both processes, cells are inoculated into a fixed culture volume and the more product needed, the larger the culture system. Cell growth is rapid in the fresh medium. As the nutrients are consumed, and waste products accumulate, growth rate declines, resulting in a culture decay phase. In a fed-batch system, the viability and productivity of the culture can be extended somewhat by supplementing it with growth promoting additives. In either case, however, declining growth and decay is a consequence of consumption of essential nutrients and accumulation of growth limiting cellular byproducts. The final productivity of such cultures may typically vary from hundreds of mg/L to a few mg/L.  

Alternatively, it is possible to extend and maintain the viability of a culture and its productive phase by continuously removing the inhibitory waste byproducts while replenishing the culture with fresh nutrient medium, in a process known as perfusion. Normally, this process is done using a filter or other cell retention device, where the protein product is collected with the waste stream while the cells remain in the reactor. Continuous removal of product through the waste stream prevents its accumulation in the vessel. Rapid removal and storage offers an important advantage with some labile products, but generally, the large volume collected is not desirable. The ATF System used in a concentrated fed-batch (CFB) process takes advantage of the high cell concentrations and resultant high levels of product formation offered by perfusion and at the same time eliminates dilution of the product. The ATF System CFB process is realized simply by using a standard molecular weight cut-off filter, typically, 50 kDa. The filter allows waste products or any compound smaller than the indicated pore size to flow through, but not larger molecules. Antibodies and many other potential proteins or products larger than 50 kDa are retained by the filter. Both cells and product can thus be concentrated to high specific concentrations. A comparison between a standard fed-batch and CFB was undertaken by Biovian to evaluate the differences between these two processes.  

: Refine Technology Alternating Tangential Flow (ATF2) System using the F2 Hollow Fiber Module 50 KDa pore, PS, 1 mm ID, 0.14 m

: Wave Cellbase 20 PS with control unit Wave Biotech BWC 

: Sartorius-Stedim Cultibag RM 2 L Optical ATF and Opta SFT sterile connector 

 CHO cell line expressing human immunoglobulin G (hIgG) 

The CHO cell seed culture was prepared in spinner flask. The cells were transferred to the wave reactor, at a starting concentration of approximately 0.4x10

  cells/mL (Day 1). The bag culture-volume was approximately 1 L. The bag was equipped with optical sensors for dissolved oxygen (DO) and pH on-line measurements. Aeration was not initiated until Day 2 in order to maintain pH. Afterwards, aeration was maintained at 100 mL/min. The oxygen concentration was maintained as instructed by the manufacturer by increasing the rock and angle parameters. 

In the fed-batch reference run, nutrients, such as CD-feed and glucose, were fed batch-wise at four different points in time during the process. The pH was automatically controlled. 

In the ATF CFB experiments, the ATF System was connected to the culture bag. The flow was started on Day 2 at a constant flow rate of 0.5 L/min. The perfusion (fresh medium in and waste medium out) was started on Day 3. In Run 1, the perfusion rate was initiated at 0.8 L/d and increased to 1.2 L/d. In Run 2, it was initiated at 0.8 L/d and increased to 2.5 L/d by Day 7. The medium addition and waste outflow rates were determined by weight change of the respective containers. 

Samples of approximately 2 mL were removed at least once a day from the cultivation bag with a syringe. DO and pH were registered during the sampling. An aliquot of the sample was immediately frozen at –20 °C for product analysis performed after the end of the actual experiment. Glucose and lactate concentrations were determined subsequent to sampling. The cell concentration and cell viability were analyzed by diluting the sample in phosphate buffered saline and tryphan blue before automatic counting of living and dead cells by a cell counter. The product was analyzed by a hIgG immunoassay. 

The results of the fed-batch culture shown in Figures 1–6 are typical of such cultures. There was rapid loss of viability following a growth phase (see Figures 1, 2), continued increase in product concentration after the culture began to decline (see figure 3) and accumulation of waste products (lactate, see Figure 4), accompanied by a steady decline in glucose concentrations and DO (see Figures 5, 6). Supplementing the fed batch culture four times with nutrient media during the course of the culture resulted in a peak cell concentration of 5.8x10

 cells/mL at 191 h (see Figure 1). As expected, the highest product concentration was somewhat delayed, reaching 0.4 g/L, at 260 h (see Figure 3).  

Figure 1: Number of viable cells in fed-batch and concentrated fed-batch runs. (ALL FIGURES ARE COURTESY OF THE AUTHORS) 

The benefits of addition of fresh medium while removing waste media is also illustrated in Figures 1–6. pH was set by the process without the need for a separate pH control (see Figure 7). The perfusion rate in this run varied between 0.8 and 1.5 vv/d (see Figure 8). The first run was less than ideal. The weight measurement for the Wave bag and associated control loop to keep the level constant in the bag had a problem resulting in a working volume that was too high. A manual adjustment was made by increasing the filtrate flow rapidly to remove this excess media.  However, this adjustment was too fast for the ATF System's cleaning action and the essentially dead-end filtration performed led to filter failure. Viability started to decline early in the run, at <150 h, and the viable cell count also leveled off at about the same time (see Figures 1,2). The perfusion was ended at 239 h and in spite of the dificulties, a peak cell concentration of 16.4x10

  cells/mL, was reached at 195 h (see Figure 1), whereas the highest product concentration, 1.9 g/L, was reached at 239 h (see Figure 3).  

Figure 2: Percent of viable cells in fed-batch and concentrated fed-batch runs. 

The second CFB run with the ATF System differed somewhat from the first CFB run. It was performed with a more rapid increase in perfusion rate to about twice the final rate of the first run. The perfusion rate in this run varied between 0.8 and 2.5 vv/d (see Figure 8). The cell growth was sustained longer than in the first run, achieving a significantly higher cell concentration. The highest cell concentration, 34.5x10

  cells/mL, was reached at 284 h (see Figure 1), whereas the highest product concentration, 3.8 g/L, was reached at 332 h (see Figure 3). The viability of the culture was sustained above 85% for the duration of the culture (see Figure 2). After an initial adjustment, glucose and lactate were sustained at about constant levels for most of the culture duration (see Figures 4,5).  

Figure 3: Product concentration in fed-batch and concentrated fed-batch runs. 

The results in Table I show that cell, and more importantly, product concentration can be greatly enhanced with the ATF System CFB process. Even in the first CFB run which was a less than optimal culture, nearly three–fold higher cell concentration was achieved and more than a 4–fold increase in product concentration was obtained than in fed-batch, in about the same time period. The results improved further in the second CFB run, increasing product concentration to nearly ten–fold. Media feed rate appears to have had a great affect on cell growth rate and product levels. Further enhancement would be possible with enrichment of oxygen transfer and media development.  

Figure 4: Lactate concentration in fed-batch and concentrated fed-batch runs. 

Despite the increases in product concentration in CFB over fed batch, the ultimate productivity and the economical aspects of both process modes need to be assessed and understood. Some considerations are reactor volume capacity, and the amount of product needed, as well as the time taken to produce the amount of product.  

Figure 5: Glucose concentration in fed-batch and concentrated fed-batch runs. 

Using the data obtained in these experiments, fed-batch is further compared to CFB. Three production scenarios, (Case A, B and C in Table II), are offered for producing 3.8 g of hIgG, which was achieved in the second ATF run, and 100 g of hIgG.

Figure 6: Dissolved oxygen (DO) in fed-batch and concentrated fed-batch runs. 

Table II can be used to evaluate requirements for production of a specified amount of product and to further analyze whether those requirements correspond to a user's needs or capabilities. For example, if the user has 1-L reactors, but wishes to produce 4 g of product, then it will require over 3 months and 9 batches to do so, using the fed-batch mode (Case B), but will only need 1 batch and two weeks in concentrated fed-batch mode (Case C). Given the risks of running 9 batches versus 1, and the 3 month extra overhead cost, there is a significant advantage in adopting concentrated fed-batch, at the cost of spending as little as 2.5 times more on media.  

Figure 7: pH in fed-batch and concentrated fed-batch runs. 

The relationship above is obviously relevant for any size reactor: If the user has a 100-L reactor, perhaps a single use bioreactor, and needs 400 g of product, it will require about 10 batches and four months with a fed batch process (Case B). The user may consider buying a new 1000-L reactor, which would do the job in one batch, but will also require associated accessories, downstream equipment and facility space. Alternately, the user may use the ATF System, which will deliver the 400 g required in one batch in the existing 100-L (Case C).  

Figure 8: Flow rate in concentrated fed-batch runs. 

If the user has three reactors, 25-L, 100-L, and a 250-L to choose from, and 100 g of product is needed, then the 250-L fed-batch option would seem to come out slightly better than the 100 L fed-batch, with a longer production cycle, or the 25-L CFB system with a similar production timeframe but costlier media use. However, when considering seed expansion for this 250-L reactor, which is not factored into the examples in Table II, it appears that the 25-L and 100-L reactors are needed anyway; therefore, when assessing risk versus the gain in using the 25-, 100- or 250-L system, it appears to be similar in each case. Other factors such as downstream processing of 25-L versus 250-L harvest need also to be taken into account.  

In yet another scenario: What if the user has several 25-L reactors and only one 250-L reactor but wishes to produce multiple products in a limited time? Clearly the concentrated fed-batch approach can most readily achieve that goal. The multiple 25-L reactors can be used simultaneously, rather than waiting for the main reactor to become free. The 25-L reactors will also not be tied up as seeds for the 250-L reactor.  

Table II: Production of human Immunoglobulin G (hIgG) using fed-batch and concentrated fed-batch. 

The use of the ATF System to achieve exceptionally high productivity when used in a concentrated fed-batch process lends itself to more efficient transition to upstream operation. Additionally, in the case of a midsize or smaller facility, the requirement of expanding capacity through reactor scale-up is removed. The ATF System allows existing equipment to be more flexibly adjusted to increased production requirements. 

Features

Cell concentrations and resulting protein concentrations are higher in a concentrated fed-batch process than in a standard fed-batch culture system.

The Use of the ATF System to Culture Chinese Hamster Ovary Cells in a Concentrated Fed-Batch System

Achieving operational excellence is a growing goal among biopharmaceutical enterprises. Companies are investing heavily in software, data marts, and data integration  to achieve operational excellence. The authors explore approaches for meeting operational-excellence goals, and discuss how to manage collected data with a risk-based approach for improved process understanding. 

Operational excellence is achieved when each and every employee can see the flow of value to the customer, and fix that flow when it breaks down (1). To reach this goal, and thereby to achieve continuous improvement at all levels of organization, one needs to make the best possible use of available resources (e.g., data, information). Only when there is complete visibility of existing manufacturing processes and systems within an organization can this goal be achieved. In the biomanufacturing industry, 

 means effective communication of process data and product information.  

Quality by design (QbD) is defined in the the International Conference on Harmonization's Q8 Guideline as "a systematic approach to development that begins with predefined objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management" (2). In a typical QbD approach for development of biological products, decision on the Target Product Profile (TPP) is followed by a risk- and science-based assessment of product attributes with respect to their effect on the product's safety and/or efficacy (3, 4). This assessment results in identification of the so-called critical quality attributes (CQAs). Next, the process is designed to consistently deliver these CQAs within the acceptable limits.  

A robust control strategy that can ensure consistent process performance in view of the various sources of variability that exist in biotechnology manufacturing is central to successful implementation of QbD. Once the manufacturing process and the control strategy are in place, validation is performed to demonstrate the effectiveness of the control strategy, and the product is filed for approval. After approval, ongoing monitoring is performed to ensure robust process performance over the life cycle of the product (2, 3).  The approaches needed to make these decisions require sufficient access to process and product information that is otherwise distributed within a myriad of sources. Thus, the need for efficient information-sharing has become more crucial to industry in the QbD paradigm. 

about the "Elements of Biopharmaceutical Production" (see full list at 

). This article focuses on operational excellence in manufacturing of biotechnology therapeutic products in the QbD paradigm, including the use of efficient and effective data and knowledge management. The authors discuss key concepts and present a related case study. 

DATA AND KNOWLEDGE MANAGEMENT: THE TRADITIONAL APPROACH

In an attempt to achieve operational excellence, biopharmaceutical companies often spend significant amounts of resources on building data-collection and data-capture systems, leaving fewer resources for the more important task of performing analysis of the collected data. Collecting more doesn't enable doing more. Rather, the task adds to the complexity of finding a needle in haystack. Although data collection is an important step toward continuous improvement, the inability to visualize data and share its analysis across the organization is a key deficiency in current industry practice. 

Complex statistical tools for performing advanced analysis of manufacturing process data have become the norm in biotechnology manufacturing (5). As the complexity of process data and analysis increases,  however, training an entire organization's staff on how to best use these tools is not feasible. Many organizations, therefore, opt to create a specialized team that is trained to perform such analysis. As a result, organizaitons end up being segregated into groups of staff who on one side, have hands-on scientific experience with the product or process but no knowledge of the analytical tools associated with them, and on the other side, staff who know how to use the statistical tools but do not have hands-on scientific experience with the product or process. This situation is not desirable.Appropriate analysis requires both sides of expertise.   

DATA AND KNOWLEDGE MANAGEMENT: THE QBD APPROACH

To make the most out of collected data and information, one must monitor, explore, and analyze it in a way which is easy to access and understand by the majority of the staff. This goal can be achieved in the following manner:  

Apply risk-management principles to data analysis. For all parameters, a company may continue to record data on paper (e.g., batch records) or in electronic format. Data archiving is a minimum expectation of cGMP manufacturing. However, more sophisticated forms of data archiving, visualization, and analysis may be reserved for parameters that have been prioritized. This point is further illustrated in the case study presented later in the article. 

Create domain-specific visualizations that can help to monitor processes and serve as a standard way of communication of process data across the organization. Visualizations can also provide a quick glance into the status of the process. The effectiveness of visualizations depends on how well they  represent the relationship between different types of data (e.g., batch performance data, time profiles, batch events). Visualizations should be accessible to all staff members associated with the process and the product. 

Perform advanced data-capture and statistical analysis to gain deeper insights into the manufacturing process. These tools should be used (or piloted) by a smaller subset of the organization but should be introduced to the broader organization only after thoughtful consideration.

The primary goal of the proposed approach is to provide access to simple, easy-to-learn tools and approaches to the majority of the company's staff. As discussed in the precious section, this approach alleviates a significant portion of the inefficiencies that are otherwise present in the traditional approaches of data and knowledge management. 

Figure 1 illustrates an implementation roadmap for a typical biomanufacturing process where data is created at multiple sources. Several hundred process parameters are collected for each batch during process execution. Because monitoring each parameter in such a large dataset will require significant resources, a more practical approach would be to use risk-assessment and management principles to prioritize the process parameters. Risk-assessment tools such as the Failure Modes and Effects Analysis (FMEA) can be used effectively for this purpose. 

In this approach, each parameter is assessed for its importance to the process. Questions that need to be asked may include: What is the severity of the impact of the parameter on the performance of the given process step and the quality of the product? Is the parameter part of the process control scheme? How often does the parameter go outside the allotted operating range? Is the parameter alarmed?  

Based on this analysis, a risk priority number (RPN) can be calculated and used to prioritize the parameters. Based on process-execution experience and current level of process understanding, a cut-off RPN number can be determined to screen only those parameters for process monitoring that are above the cut-off RPN (see Figure 1). The data about parameters below the RPN cut-off can be archived for later use in compliance and advanced analysis purposes. The parameters identified for routine process monitoring can be represented in simple visualizations that slice and dice data in various combinations of batches, parameters, and process time (e.g., parameter versus time, parameter versus parameter, parameter versus batch). These visuals can help the organization spot process trends and communicate and resolve issues easily. Unresolved issues that need a deeper understanding of the process can be resolved by using advanced statistical analysis tools. The ideal outcome of advanced analysis is process improvement initiatives that are aimed to improve product yield and quality (6), as well as process-analytical-technology (PAT) based control schemes aimed at ensuring consistency in the product yield and quality.  

Figure 1: Illustration of a quality-by-design approach to data and knowledge management. (ALL FIGURES ARE COURTESY OF THE AUTHORS) 

CASE STUDY: A QUALITY-BY-DESIGN APPROACH FOR A MAMMALIAN CELL-CULTURE STEP

Figure 2 lists typical input and output parameters that are encountered in routine mamxmalian cell-culture applications. These data may come from online sensors that are collecting the data in real time, from batch records, from analysis in the Quality Control laboratory, or from the various benchtop instrumentation that accompanies the bioreactor. According to cGMP expectations, these data are collected and archived. Performing a statistical analysis of each parameter would be cumbersome and unlikely to add much value with respect to process understanding. An FMEA, therefore, was performed to segregate the parameters into multiple categories (see Figure 3).  

Figure 2: The various input and output parameters typically encountered when performing mammalian cell-culture processes. DO is dissolved oxygen. RPM is the agitation rate. RVLP is Rubella virus-like particles. MMV is mouse minute virus. 

Category A lists parameters that are collected and archived for future potential use. Category B lists the parameters that have been identified as important for the process step. The data are routinely monitored, which involves performing statistical analysis of the data and trending. Analysis 

 Nelson rules is a popular approach for performing this activity (6).  

Figure 3: Outcome of risk analysis for prioritization of various input and output parameters typically encountered when performing mammalian cell-culture processes. Data for parameters are collected and archived. Only a subset of parameters are routinely monitored and a further smaller subset is involved in process-analytical-technology (PAT) based control of the process step. 

Finally, a subset of Category B parameters may be used for designing a PAT-based control scheme. For example, a PAT control scheme for gas transfer would need measurements of pCO

, agitation rate (RPM), and air-flow rate (Category D). Hence, these parameters may be collected using automatic sensors and the information relayed to the controller. Similarly, a PAT control scheme for cell count will require efficient capture of information on pH, temperature, cell count, agitation rate, air-flow rate, and feed-flow rate.  

This article aims to illustrate the need to achieve operational excellence with respect to data- and knowledge-management systems to successfully implement QbD for manufacturing of biotherapeutics. As discussed, there is a great need among the biomanuacturing industry to apply risk-assessment and management principles to achieve this objective. When done correctly, this approach is more economical and effective than traditional approaches otherwise used in industry. 

are cofounders of Simplyfeye Softwares in Delhi, India. 

is a partner at Simplyfeye and a faculty member at the Indian Institute of Delhi, India, +91-9650770650, 

. Rathore is also a member of BioPharm International's editorial advisory board.   

 1. "Operational excellence" as defined by 

www.industryweek.com/articles/operational_excellence_defined_14011.aspx

 6. P. Konold, R. Woolfenden II, Cenk Undey, and A. S. Rathore, 

The authors focus on operational excellence in manufacturing of biotechnology therapeutic products in the QbD paradigm.

Operational Excellence: More Data or Smarter Approach?

The use of single-use systems has become one of the biggest trends in biopharmaceutical manufacturing since the introduction of complex and sensitive biomolecules, such as monoclonal antibodies and recombinant protein biotherapeutics. In particular, single-use technologies have been widely adopted by global vaccine manufacturers as a means to eliminate cleaning and cleaning validation issues following the influenza vaccine shortage of 2005, which was attributed to contamination issues in a production facility located in the United Kingdom.  

There has already been much discussion on single-use systems at US conferences and in US publications during the past 10–15 years, but interest is now expanding further across the globe to Europe and Asia. Despite this growth, there have been only a few dedicated conferences in these regions that focus on single-use systems, even though significant implementation in European vaccine and biopharmaceutical manufacturing facilities has occurred. However, this trend may be changing. This year, I've been privileged to speak at and chair several international conferences focusing on single use. Such conferences provide an important opportunity for industry participants to share experience, concerns, and best practices.  

On February 1–2, 2011, I visited Brussels to chair the International Quality and Productivity Center's (IQPC) Pharma–IQ Disposables Solutions for Biomanufacturing Conference, which was attended by nearly 100 European manufacturers and suppliers. The conference was Pharma–IQ's third Annual European Disposables meeting. The previous two annual conferences were held in London, and the event is shaping up to be a key European conference for single-use activities. This year's meeting featured speakers from several European-based vaccine, biopharmaceutical, and contract manufacturers, including Novartis, Vivalis, Genzyme, CMC Biologics, Cobra Biomanufacturing, Innogenics, Crucell, Eli Lilly, GlaxoSmithKline, Pfizer, and Merck Serono, along with supplier presentations from Pall Life Sciences, Sartorius-Stedim Biotech, Novasep, JM Bioconnect, GE Healthcare, and ATMI Life Sciences. 

National European regulatory authorities and the European Medicines Agency have not made public statements about the acceptability or requirements for single-use technologies, but this has not deterred European drug sponsor and contract manufacturing facilities from implementing such systems for both clinical and production batches. Conference topics, for example, focused on everything from comparability of product quality and reduction of carbon footprints to implementation of single-use systems for buffer filling processes and preparation of regulatory submissions. The conference also featured several roundtable discussion sessions that facilitated the sharing of questions and experiences among participants.  

Shortly after Pharma–IQ, I visited Tokyo (just before the earthquake and tsunami occurred) to give a presentation on single-use technologies and quality requirements at the International Society for Pharmaceutical Engineering (ISPE) Japan Sterile Products Processing Community of Practice meeting. Compared with Europe and the US, implementation of single-use systems in Japan has not been as extensive. Japanese pharmaceutical companies, however, are interested in new applications for single-use technologies. I also met with representatives from Japan's Ministry of Health, Labor and Welfare, who were eager to learn about best practices for implementing single-use systems in biopharmaceutical manufacturing. 

At the end of February, I attended the ISPE Tampa Conference in Florida, where I spoke at an aseptic processing seminar about sterilizing filtration. One of the hot topics discussed is related to European GMP Annex 1 recommendations, which call for integrity testing of sterilized filters prior to use and double filtration schemes to enhance sterilization assurance. These recommendations can lead to complex systems and procedures, but are possible when using properly designed single-use systems (as opposed to stainless steel systems). One concern, however, is that the EMA cites a potential subtle change in membrane retention properties that might occur with exposure to moist heat (steam sterilization), but could also potentially be resolved by filter plugging during the process.   

With single-use filtration systems, sterilization is most commonly performed with gamma irradiation, which does not introduce heat and has not been associated with changes to membrane filter porosity or bacterial retention property (so long as the filter materials are gamma compatible and validated for gamma irradiation stability). Single-use filtration systems are typically supplied presterilized, so that a poststerilization integrity test is the only pre-use test a user can apply. Even without the hypothesized potential changes in membrane flow path and retention property caused by moist-heat sterilization conditions, it can still be economically prudent to conduct a pre-use (poststerilization) integrity test in many single-use filtration processes. This step ensures that the filter has not been damaged during shipment and handling. Such damage is not repairable by plugging and is a more valid reason to conduct pre-use integrity testing of the sterilized filter, especially where a drug product is sterile filtered through single (nonredundant) filtration schemes and filled into multiple final containers, or that cannot otherwise be refiltered in the event of post-use filter integrity failure. When designing single-use single or double filtration systems, it is important to consider how the filters will be initially wetted, how they will be tested, and whether or how to displace the wetting liquid before product introduction. Approved system design should be discussed with the filter supplier, system-design engineer, and system integrator.  

In March, I finished my single-use tour by chairing the second annual IBC Biomanufacturing and SingleUse Systems Asia Conference, which took place in Shanghai. Last year, the event was held in Seoul, South Korea. I was immediately struck by the progress in appreciation for the benefits of single-use manufacturing by many of the Chinese participants. Single-use manufacturing has been a difficult concept for many Chinese biopharmaceutical manufacturers to accept because they have historically been focused on minimizing cost per batch, rather than the degree of cleaning and cleaning validation required in the West, or other beneficial aspects of single-use systems, such as reduced carbon footprint. At this meeting, participants heard case studies and considerations for single-use technologies from companies across Asia, including Simcere Pharmaceuticals, Shanghai Kanda Biotech, Wison, and Autekbio, all based in China; Novartis and Green Cross, based in Korea; Innobioventures, based in Malaysia; as well as from Piramal Healthcare and Intas Biopharmaceuticals, both based in India. In Asia, much of the biopharmceutical focus is on the development of biosimilars. Although the cost of single-use technologies versus cleanable and reusable stainless steel is still an issue for many process developers in Asia, disposable bioreactors and single-use tangential flow filtration are now being seen as potential ways to accelerate biosimilar development. 

For those involved in or considering single-use processes, two upcoming US conferences may be of interest. The PDA Single-Use Workshop, taking place June 22–23 in Bethesda, Maryland, will include details about PDA's technical report on single-use technologies using quality-by-design principles. Workshop attendees will have the opportunity to preview and discuss the draft report during the workshop so that industry input can be incorporated into the final version, which will be published by PDA in fall 2011. See 

The Bio-Process Systems Alliance International Single-use Summit, to be held July 27–30 in Washington, DC, is meant for biopharmaceutical business leaders and will focus on the impact of single-use manufacturing on the future of healthcare and biopharmaceuticals. The meeting will feature speakers from PricewaterhouseCoopers, Venable LLP, and FDA, as well as international single-use end-users and leading suppliers. See 

is senior vice-president of Global Scientific  Affairs at Pall Life Sciences, Port Washington, NY, and chairman of the Board and Technology Committee at Bio-Process Systems Alliance, tel. 516.801.9086, 

Disposables Advisor

Single-Use Technologies World Tour

 adventitious virus screening of a bulk harvest from a mammalian cell biopharmaceutical manufacturing process, the authors obtained results that at first suggested a viral contaminant, but that subsequently were found to represent a contamination with a nontuberculosis mycobacterium (

Eukaryotic cell processes used in the manufacture of biologics are susceptible to contamination with adventitious agents. These agents include viruses, bacteria, fungi, mycoplasmas, and the so-called infectious proteins, such as transmissible spongiform encephalopathy agents. These entities may be introduced through contaminated raw materials (reagents and cell lines), during cell culture scale-up and fermentation, or during downstream processing. The methods used to conduct safety testing of biologics produced in cell cultures, and a survey of the types of contaminants that have been detected, have been described previously (1). The authors describe contamination of a mammalian cell-based biologic manufacturing process with a nontuberculosis mycobacterium (

A bulk harvest from a mammalian cell bioreactor process was submitted to the authors for evaluation in a 28-day 

 adventitious virus screening assay as part of an investigation of a bioreactor problem at a biologics manufacturing site. The bioreactor run had been terminated early, because of an abnormally high value for one of the in-process monitoring parameters. Microbial testing of the bioreactor contents, performed at the manufacturing site, had indicated an acceptable level of bioburden. As part of the investigation of the failed bioreactor run, the authors assayed the sample from the terminated bioreactor for potential viral contaminants according to  the routine screening assay protocol. Eight days after inoculating the clarified bulk harvest material onto human (MRC–5), African green monkey (Vero), and Chinese hamster ovary (CHO–K1) detector cells, we observed floating clumps of an undetermined nature in the growth medium covering the monolayers. By 14 days post inoculation, the MRC–5 culture was devoid of viable cells. The Vero and CHO–K1 monolayers were intact and fully confluent at this time, though each displayed the floating clumps. Hemadsorption and hemagglutination testing of the various cultures, using chicken, guinea pig, and rhesus monkey erythrocytes, was completed on day 14 with negative results. The authors performed a passage of conditioned medium from each day-14 culture onto fresh cultures of detector cells. Eight days later, the MRC–5 monolayers displayed reduced confluency and abnormal cell morphology, including vacuolization. The Vero and CHO–K1 monolayers remained fully confluent at this time, although each displayed the floating clumps noted above. The hemadsorption and hemagglutination results were again negative. The MRC–5 portion of the study was repeated in its entirety from the beginning, and similar results were obtained. 

Conditioned medium samples harvested from the MRC–5 cultures displaying deterioration were filtered (0.45 μm pore size), and the filtrate inoculated onto fresh cultures of MRC–5 cells. When the secondary MRC–5 cultures were observed for 14 days, the authors did not see a reappearance of the floating clumps or the progressive cell deterioration. This procedure was repeated, with similar results being obtained. In contrast, passage of the same conditioned medium samples (albeit without filtration) onto fresh MRC-5 cultures resulted in the reappearance of the floating clumps and progressive cell deterioration within a few days.  

The filtration results described above indicated a microbial, not a viral, origin for the effects noted. The authors subsequently verified this idea through transmission electron microscopic examination of the affected MRC–5 cells, which revealed organisms both within (see Figure 1) and outside of the cells (see Figure 2). These organisms were rod-shaped, with diameters ranging from 0.34 to 0.39 μm and overall lengths of up to 1.1 μm. The organisms were not apparent on visual observation of the MRC–5 cell monolayers, using light microscopy. They were, however, readily observed when the focal plane was raised to that of the medium–air interface. At this level, which is not typically viewed when scoring cell cultures for cytopathic effect, the authors could easily visualize both the floating clumps noted at the monolayer and mats of dark, branching organisms (see Figure 3).  

Figure 1: Transmission electron micrograph of an MRCâ5 cell from an infected culture, showing intracellular organisms (B), 36700 x magnification. (ALL FIGURES ARE COURTESY OF THE AUTHORS) 

A Sabouraud dextrose agar (SAB) plate streaked with conditioned medium samples from the affected MRC–5 plates and incubated aerobically at 20–25 °C led to white- to cream-colored colonies by day 7. The organism also was isolated using fluid thioglycollate medium incubated aerobically at 30–35 °C. In the latter case, growth was observed by day 7 as particulates that were found in a band about two thirds of the distance to the top of the liquid, and in a band floating at the liquid/air interface. Isolation of the organisms on tryptic soy agar (TSA) plates resulted in white- to cream-colored colonies by day 7 after streaking of conditioned medium samples from the affected MRC–5 plates. These colonies were present both on TSA plates incubated aerobically at 20–25 °C and in plates incubated at 30–35 °C. Gram staining of the organisms revealed a weak, inconsistent positive reaction. The organisms gave a positive reaction, staining as dark red and pink rods, when evaluated using Kinyoun's acid-fast staining procedure. Colonies taken from the TSA plates were submitted to a subcontractor for identification. Using the 16s rRNA gene sequencing method, the isolate was found to be a member of the 

 complex. Differentiation of members of this nontuberculosis complex (

 was obtained, and definitive identification to the species level was not pursued.  

Figure 2: Transmission electron micrograph of an MRCâ5 cell from an infected culture, showing extracellular organisms (B), 23600 x magnification. 

The authors subsequently evaluated additional retained bioreactor harvest samples that had been collected at various intervals to establish whether the microbe was present in the bioreactor before termination of the run. The earliest bioreactor harvests did not cause MRC–5 cell deterioration, and organisms were not detected in the inoculated detector cell cultures. In contrast, all samples from a point approximately midway through the run onward elicited these effects. The latency of onset of the observed effects in the detector cell cultures correlated generally with bioreactor harvest day, such that the most rapid effects were observed in the samples harvested just before termination of the bioreactor run. Isolation of organisms from the affected studies led, in each case, to identification of the 

Figure 3: Light micrograph of an infected MRC-5 culture, showing mats of organisms and floating clumps at the plane of the medium/air interface (50 x magnification). 

The nontuberculosis mycobacteria appear to represent a potential risk to the biopharmaceutical industry, due to their propensity for forming biofilms, their ability to proliferate in water under relatively low nutrient conditions, their resistance to typical water disinfection methods, and their relatively slow growth in nutrient media (2–5). The former properties render these organisms capable of existing in water piping and other surfaces in contact with water or aqueous media. Their growth characteristics make these agents easy to overlook, especially when surveillance methods, such as short-term bioburden assays, are employed. In this case study, the bioreactor run was terminated early because of an overly high value for an in-process monitoring parameter. However, routine bioburden testing performed on bulk harvest samples taken from the terminated bioreactor failed to detect the presence of the mycobacterial contaminant. Eventual detection of the contaminant in the course of adventitious virus testing of this bioreactor harvest sample was possible because that assay was relatively long in duration (28 days) and the antibiotics used in the assay (gentamicin, amphotericin B) were apparently ineffective against the species of mycobacterium involved. The route of entry of the mycobacterium into the bioreactor process was not determined.  

The mycobacteria involved in this case are pathogenic in humans, especially those who are immunocompromised (6, 7). It is difficult to estimate the frequency of occurrence of mycobacterial contamination in the biologics manufacturing industry, because some episodes may lead to premature bioreactor termination, such as that occurring in this case, with little evidence to implicate a mycobacterium. A few cases of contaminated vaccines and tissue extracts have been reported in the literature (8, 9). One can only speculate about the actual risk to the biopharmaceutical industry of this type of contamination. 

, was a scientist, all at Biologics Testing at BioReliance, Rockville, MD,  

 1. R. Nims et. al., "Adventitious Agents: Concerns and Testing for Biopharmaceuticals,"  in: 

Process Validation in Manufacturing of Biopharmaceuticals: Guidelines, Current Practices, and Industrial Case Studies,

 A.S. Rathore and G. Sofer, Eds., (CRC Press, Boca Raton, FL, 2005), pp. 143–167. 

 3. L. Hall-Stoodley, C.W. Keevil, and H.M. Lappin-Scott, 

 6. B.G. Metchock et. al., "Mycobacterium," in: 

 Murray, et al., Eds., (ASM Press, 7th ed., 1999) pp. 399–437. 

 Raymond Nims is currently at RMC Pharmaceutical Solutions, Inc., Longmont, CO, Alfred Chun is currently at Cangene bioPharma, Baltimore, MD, and Suzanne Dieringer-Boyer is currently at the National Biodefense Analysis and Countermeasures Center (NBACC), Frederick, MD.  

The authors present a case study identifying a contaminant.

Nontuberculosis Mycobacterium Contamination of a Mammalian Cell Bioreactor Process

Large-scale biopharmaceutical manufacturing equipment for upstream product manufacture has changed relatively little over the past decades. Innovation in bioprocessing equipment, particularly, bioreactors, has been slow despite some gains, such as cost improvements, greater flexibility, higher product yields resulting from improved genetic and cell engineering, and widespread adoption of small-scale, single-use bioprocessing equipment.  

Data from BioPlan Associates' 8th Annual Report and Survey of Biopharmaceutical Manufacturing Capacity and Production show that the industry is demanding better upstream and downstream processing. The survey, which had responses from 352 global biomanufacturers, analyzed production, budgetary, technological, and quality issues. The study also showed that the industry is interested in new-product innovation in single-use bioreactors, cell-culture media, and various service areas (1). 

A key finding of the study was interest in upstream equipment innovation. Bioreactors are one of a company's largest bioprocessing expenses. To illustrate interest in alternative bioreactors, more than two-thirds (68.1%) of survey respondents reported current in-house use of single-use bioreactors, and this percentage is growing, especially in research and development and clinical-scale production. Improvements in bioreactors, particularly for single-use, along with purification equipment, were ranked as the areas where bioprocessing innovations were reported as most needed, with 29.2% of respondents citing bioreactors as a key area. Survey respondents also cited the need for improvements in cell-culture media (29.2%), upstream process-development services (18.7%), cell-line services (17.8%), and process-development services for formulation (11.0%) (1).  

Partly due to the slow-changing and strict regulatory environment, single-use suppliers are focusing on relatively traditional bag-liner-in-a-steel-bioreactor single-use systems. Much of industry's desire for improved bioreactors is related to the need for large-scale (e.g., >1000 L batch-fed), single-use/disposable bioreactors. Today, 1000 L is the largest cost-effective size for disposable batch-fed bioreactors. Future areas for single-use bioreactor innovations may include unitary (single-piece) all-plastic rather than plastic-lined and even stainless steel-lined plastic bioreactors. 

Contract manufacturers have more interest in improved single-use bioreactor systems compared with product developer/manufacturers, according to the study. Interest in disposable bioreactors also differed among regions. Respondents from Europe (32.2%) showed the highest interest compared with 26.9% from the United States, and 31% from the rest of the world (1).   

The study also surveyed 175 suppliers and examined 52 equipment and functional areas. The results showed most suppliers are involved in new-product development and that single-use bioreactors are the hottest items in new-product development; 40.5% of respondents are working on them in some way. Expression-system platforms and cell-culture optimization also were key areas of new-product development (1).  

The study further evaluated 15 areas where biomanufacturers were actively implementing performance improvements in their facilities. The greatest percentage of facilities had "significant" or "some" improvement from better process development (identified by 66.8% of respondents) and optimized cell-culture processes (identified by 61.9% of respondents) (1).  

Perfusion bioreactors appear to be positioned to become the next major revolution in bioprocessing and single-use equipment, with perfusion equipment on track to compete head-to-head with conventional batch-fed bioreactors even at commercial GMP scale. The study, however, showed that the industry's view of perfusion largely is dated.  

Perfusion is not new. For example, perfusion bioreactors were widely used for hybridoma culture in the 1970s–1980s before antibody manufacture switched to batch-fed mammalian cell bioreactor manufacture of recombinant antibodies. Perfusion bioreactors were available from such companies as Endotronics, Cellex, Biosyn, and Biovest. Today, providers of perfusion bioreactors include FiberCell Systems and Zellwerk (marketed in the US by Glen Mills). 

Nearly all current bioprocessing today involves batch-fed bioreactors. Perfusion involves continuous slow feeding and removal of spent media along with the desired product with the host cells retained within the bioreactor by being bound to bundles of capillary fibers or other membranes or retained in suspension in the bioreactor by special filters. While batch-fed bioreactors involve cells in dilute culture media suspension, perfusion bioreactors grow cells at ≥100 to 1000 times higher concentration, with bioreactors commensurately smaller. Thus, using a small perfusion bioreactor over time with the same amount of culture media may match or beat the product output of a much larger batch-fed bioreactor. For example, FiberCell Systems reports it is developing desk-sized units that will have the equivalent recombinant-protein production capacity of conventional 2000–5000-L bioreactors.  Also, ATMI recently launched iCellis, the first large-scale perfusion bioreactor. 

Although perfusion appears to be a viable new product candidate, the BioPlan survey shows that industry knowledge and attitudes toward perfusion remain largely dated. Among a list of 17 problems commonly encountered in bioprocessing, respondents consistently had more serious concerns with perfusion than with batch-fed systems. These issues included process complexity (66.1% of respondents had serious concerns with perfusion systems compared with 4.9% for batch-fed) and contamination risks (63.9% had concerns with perfusion compared with 3.8% for batch-fed systems) (1). These and other concerns for perfusion already are being resolved as new bioreactors enter the market. As new products are launched in coming years, commercial-scale devices are likely to capture significant interest as new upstream solutions. 

is president of BioPlan Associates, tel. 301.921.5979, 

 and a periodic contributor to Perspectives on Outsourcing.  

 1. E.A. Langer, 8th Annual Report and Survey of Biopharmaceutical Manufacturing Capacity and Production,  BioPlan Associates (Rockville, MD, April 2011), 

Perspectives on Outsourcing

A recent industry survey shows keen interest in improving bioreactors and cell-culture media.

Better Upstream Technologies

The proper production of safe and effective prescription drugs is central to a number of important regulatory and public policy issues now in the spotlight. The White House is orchestrating a campaign to reduce abuse and misuse of prescription drugs, with a focus on extended-release opioids. The spread of adulterated drugs across the globe is prompting initiatives to curb violative imports. At the same time, patient groups and physicians are anxious to resolve shortages of important therapies and ensure access to needed medicines. FDA is a prominent player in the crackdown on opioid abuse and oversight of drug imports, and is also keeping an eye on possible disruptions in supplies for patients. These developments add to pressure on manufacturers to secure supply chains, comply with GMPs, and to develop products that resist tampering and abuse.  

From high-security prisons to cancer clinics, crucial drug shortages are placing attention on drug manufacturing and supply issues. Disruptions in access to life-saving chemotherapies, anesthetics, and pain medications have made headlines, along with difficulties for law enforcement officials seeking approved drugs for lethal injection. Many of the shortages involve generic sterile injectibles that have been hit by a number of problems: industry consolidation that reduces competitive production, limited supplies of active ingredients, and manufacturing difficulties that have led to plant closures. Generic-drug manufacturer Teva, for example, had to shut down a California injectable drug facility after an FDA inspection cited numerous GMP violations. Compounding the difficulties is the increased import of active ingredients from foreign suppliers, China in particular. 

A March 2011 survey by the Premiere Healthcare Alliance found that last year, more than 240 drugs were either in short supply or unavailable in the US. Efforts to obtain alternative therapies boosted hospital and provider costs by at least $200 million. Even worse, shifts to different medicines appear to boost medication errors as practitioners make mistakes mixing and administering less familiar products.  

In response, FDA has moved to approve the import of cancer and leukemia therapies produced overseas. The agency also has authorized prison officials to import sodium thiopental from unapproved foreign producers, a move that generated lawsuits arguing a lack of evidence for potency and efficacy of the imported drug, along with opposition from foreign governments opposed to using their products for capital punishment.   

The situation has prompted calls to bolster FDA authority to require advance notice from more manufacturers about plans to discontinue production of short-supply drugs. The agency has fairly limited notification authority and often receives scant information from manufacturers on factors causing new shortages. Pharmaceutical companies generally oppose long advance notification requirements, noting that supply changes often cannot be predicted far in advance, and that manufacturing problems usually occur without warning.   

While some FDA offices seek to accelerate access to vital medicines made abroad, agency field inspectors are struggling to monitor a surge in imports of food and medical products. FDA has been working for more than five years to install an automated Predictive Risk-Based Evaluation for Dynamic Import Compliance Targeting computer system, known as PREDICT, to replace its outdated import screening process, which cannot handle the approximately 24 million shipments expected to arrive at US ports of entry this year. The import volume includes up to 40% of drug products and 80% of active pharmaceutical ingredients.  

The new system aims to prevent future heparin crises by shifting from inspecting imports at point of entry to an approach that reaches "back further to where products are manufactured" to ensure that drugs meet quality standards beginning where they are produced, explained FDA Commissioner Margaret Hamburg at an April 2011 hearing before the US House Energy and Commerce Committee. FDA critics complained, however, that PREDICT is deployed in only 4 of 20 FDA district offices and pressed Hamburg to do a better job. The commissioner blamed delays on software problems and noted that PREDICT was being expanded from California, Seattle, and New York to field offices in Florida and San Juan, Puerto Rico, bringing coverage up to 50% of all imports this year.  

The agency would gain more tools for managing imports, moreover, from proposed drug-safety legislation that authorizes FDA  to detain and destroy violative imports, to impose stiffer criminal and civil penalties on rule-breaking suppliers, and to refuse admission to products from plants that block FDA inspections. The bill, sponsored by Rep. John Dingell (D-MI), also would give FDA authority to issue subpoenas related to drugs, as with recently approved food-safety legislation, and would authorize more FDA overseas inspections, recalls of tainted products, and the collection of more manufacturing information.  

In the US, the most alarming drug-safety issue involves overuse, misuse, and abuse of extended-release opioid drugs. FDA and manufacturers have been struggling to devise a Risk Evaluation and Mitigation Strategy (REMS) since 2009 to promote safe use of these medications. The task is complicated, however, by the use of these powerful painkillers by millions of individuals suffering acute pain, which generates opposition from patient advocates and physicians to any policies likely to limit access to these important therapies.  

Yet, prescription drug abuse has become the nation's deadliest drug problem. FDA estimates that more than 33 million Americans misused extended-release or long-acting opioids in 2007, up from 29 million five years earlier. The Centers for Disease Control and Prevention (CDC) documents a rise in deaths and emergency-room visits related to prescription drug use, and unintentional overdose deaths involving opioids nearly quadrupled from about 3000 in 1999 to more than 11,000 in 2007.  

Drug-treatment programs are seeing more young patients coming in either addicted to opioid prescription drugs or in the first years of heroin use that started with opioids, explains Deni Carise, chief clinical officer at Phoenix House, the leading national drug-abuse treatment provider. "People think that taking a prescription drug is okay, and the habits quickly get very expensive," she pointed out.  

FDA unveiled its opioid REMS in April 2011 as a central component to a federal campaign to reduce prescription-drug diversion and abuse by 15% in five years. The REMS requires manufacturers to develop educational materials that will make physicians and consumers more aware of the dangers of opioids, as well as their appropriate use in treating pain. The broader program, which is led by White House Office of National Drug Control director Gil Kerlikowske, also calls for more extensive monitoring of drug use by the CDC, and for the Drug Enforcement Agency (DEA) to crack down on "pill mills" and clinics offering easy access to pain medication. Because many adolescents initially obtain these drugs from family medicine cabinets or from friends, more federal and state "take-back" events will promote proper disposal of unused or expired medications. Also, the plan supports state prescription drug monitoring programs with databases to track prescriptions for certain drugs and inform physicians and pharmacies of excessive patient drug use.  

To better educate prescribers in the appropriate use of these medications and to alert patients to dangers of drug misuse, FDA wants generic and brand opioid manufacturers to collectively design training materials for use by continuing medical educational (CME) programs, which agency officials will scrutinize to ensure objectivity, accuracy, and absence of promotional bias. The industry group also will develop common elements for Medication Guides that pharmacists will give patients.  

Elements to Assure Safe Use (ETASU) in the REMS will involve a collective strategy for evaluating the effectiveness of educational and monitoring efforts in six months, 12 months, and annually thereafter. The assessment will report on how many prescribers complete educational programs and if the REMS has any impact on patient access to opioids—for both appropriate and inappropriate uses. FDA has required similar REMS for two recently approved pain-relief products (Covidien's Exalgo and Purdue Pharma's OxyContin), and these would be incorporated into the new class-wide REMS.   

Although FDA advisory committees rejected such a voluntary educational campaign last July as too weak to curb abuse, the agency is adopting this moderate approach as a first step in trying to tackle the problem. Janet Woodcock, director of the FDA Center for Drug Evaluation and Research (CDER), noted that the agency can require drug manufacturers to develop educational materials, but lacks the authority to control physician prescribing. A limited distribution system for opioids with mandated physician and patient registration would create an oversight responsibility far beyond FDA's capabilities and generate fierce opposition from the medical community. 

Instead, the Obama administration is asking Congress to enact legislation that mandates training on opioid use for physicians to obtain the DEA registration number needed to prescribe opioids and other controlled substances. But for now, the educational component of the national plan is optional for the 600,000 doctors and other health professionals licensed by DEA to prescribe controlled substances. In 2000, Congress approved the Drug Addiction Treatment Act, which requires 8 hours of special training for physicians to prescribe and dispense buprenorphine (Subutex and Suboxone) for treating opioid addiction, but this involves much more limited drug use and far fewer physicians. The prospect of additional Congressional action in the near future is fairly dim because both the medical pain-management community and pharmaceutical industry oppose mandatory training or other initiatives that could limit access to pain medications.  

At the same time, drug-abuse groups would like to expand the scope of the REMS to include immediate-release pain medications, such as short-acting Vicodin (hydrocodone), which is the most widely prescribed drug in the US (131 million prescriptions were written for Vicodin in 2010, according to IMS Health). The current program is limited to extended-release opioids, such as hydromorphone, morphine, oxycodone, oxymorphone, and tapentadol, because these drugs contain a stronger dose of active ingredient and, thus, are more dangerous when abused.  

Just how some 16 brand and generic-drug firms will devise a singled shared system for educating prescribers and patients and for monitoring the program's impact is unclear. While components of the educational materials and Medication Guides will apply to all drugs in the class, each manufacturer also will have to provide drug-specific information. A common approach for assessing the effectiveness of the REMS will involve independent audits of training quality, evaluation of providers' understanding of opioid risks, and a broader surveillance plan to detect misuse, abuse, overdose, addiction, and death resulting from specific drugs.  

CDER's Office of Surveillance and Epidemiology may play a role by tapping data bases on prescribing, emergency room visits and other indicators that could help evaluate the program's impact. In the end, all affected manufacturers may submit a similar REMS to FDA, with appendices on their specific drug. FDA wants to receive the industry REMS plan by fall 2011 so that the program can go into effect in early 2012.  

The ultimate solution to the opioid misuse problem is for manufacturers to formulate pain killers more resistant to abuse. The White House plan calls on FDA and the National Institute on Drug Abuse (NIDA) to collaborate on research for developing and testing abuse-deterrent formulations, as well as pain therapies less attractive to nonmedical users. FDA also is charged with issuing guidance in two years on developing and testing such products and on ways to assess performance in the real world.  

Clearer standards for studying tamper-resistance and for conducting clinical trials would be useful to manufacturers developing medicines with anti-abuse features, according to the Pharmaceutical Research and Manufacturers of America (PhRMA), which testified before an April  2011 Energy & Commerce subcommittee hearing on combating prescription drug abuse. PhRMA also noted the importance of obtaining adequate reimbursement and coverage for addiction treatments.  

In fact, formulating abuse-resistant products can be challenging. King Pharmaceuticals (now part of Pfizer) had to recall painkiller Embeda (morphine/naltrexone) several times after it was approved in 2009 due to stability and dissolution problems related to its anti-abuse formulation; apparently it may not return to the market. Purdue Pharma's extended-release Palladone (hydromorphone) was associated with serious adverse events and pulled in 2005. Pain Therapeutics is developing abuse resistant Remoxy as a substitute for oxycodone. To demonstrate that its unattractive taste and texture will discourage abuse, the company designed a clinical study to document lower "drug liking" by potential abusers, which it hopes will pass muster with FDA. 

Washington editor, Chevy Chase, MD, 301.656.4634, 

Regulatory Beat

Industry struggles to curb drug abuse, diversion, and disruptions in supply to ensure access to quality products.

Safety Concerns and Shortages Challenge Pharma Manufacturing

The US Patent and Trademark Office (USPTO) is underutilized as a resource of scientific and technical literature. More than 7.7 million US patents are available for free online, at 

 for the European Patent Office). All too often, scientists show up at meetings to present posters on work that has already been invented and published by others, but because those scientists searched only the literature published in scientific journals, important science and information was lost to them.  

There is a one-year timeframe during which a publicly disclosed (as in a scientific journal or poster presentation) scientific or technical invention can still be submitted as a patent application in the United States. There is no such delay in most of the rest of the world where public disclosure precludes submission of a patent application. Understanding this restriction is important to every science and technology based researcher for two reasons.  

First, there is value to scientists (both established and students) in the potential funding of research and development (R&D) projects by private industry, which spends billions of dollars on such investments annually. Funding from federal and state governments is modest in comparison. The basic science that is available in the literature and patent databases can be turned into applied science with great benefits to society. 

Academic institutions, therefore, need to recognize the value of intellectual property in the form of patents to their externally funded research programs, and to their faculty and students (who will be searching for positions and careers outside of academia following graduation). They can do this by encouraging less traditional grant applications, such as those from industry. The National Institutes of Health is not the only grant provider to science researchers in the US.  

Second, although patents and their corresponding applications are not intended to convey the same content as peer-reviewed publications, there is vast data and knowledge included in patents. Inventors are obliged to convey the best method(s) of practicing their inventions and of providing data that supports the invention and its associated claims. Researchers can use this information to their advantage. 

There is typically a three-year wait from the time an applicant files a patent submission and the time the patent information is uploaded to the national patent database. Patent applications, however, are typically published within 18 months of filing. Although the claims in a patent application may change before a final patent is issued, the specifics of the data and ideas rarely change. Of importance to the researcher is, therefore, the availability of new information in the patent literature at an earlier time than it would appear in the scientific literature.  

The USPTO database is free and includes all patents published since 1976. The database is  searchable electronically by keywords, including names, assignees, and subject areas. A complementary tool is the PAIR system on the USPTO website, which includes documents associated with the prosecution of a particular patent application, and thus, additional useful information.  

Adding visits to the US Patent and Trademark website, in addition to reading published scientific literature, will open up new vistas to researchers as well as individuals charged with guiding technology development and innovation regardless of whether that individual works in an academic environment or in private industry. USPTO represents a tremendous resource for innovators and deserves attention. 

 is managing partner of BioScience Consultants, Norfolk, VA, 

Final Word

Patent databases are often overlooked as a source of basic science, research, and development

Free Startup Research

We use "Top 10" lists to help put order to our environment, but until recently, little effort was given to defining who is manufacturing biologics, and where. This, in our opinion, is more important to us individually than lists of companies based on revenue. While revenue analysis can define a company's distribution, marketing, and business development prowess, it does not necessarily define its core operations and manufacturing competence. Revenue reported from "headquarters" can be moved around, but manufacturing is a fixed, long-term commitment to a community, an employment base, and a strategy. 

In today's global biomanufacturing environment, buyouts, layoffs, and divestitures can take place rapidly. But establishing a manufacturing base requires long-term commitment to communities, to internal strategy, and to individual employees. 

The success of a biomanufacturing enterprise depends as much on its human capital as its fixed capital, and a critical mass of appropriately educated and skilled people is often needed to establish a viable biotech cluster. Funding alone isn't enough to build a cluster. Neither is raw access to life-scientist communities (for example, the 9000+ scientists around the National Institutes of Health and FDA hasn't produced a Top Biotech Manufacturing Cluster for the Maryland or Washington DC area). A successful cluster requires the right mix of people and funding in addition to other factors, as our research at BioPlan Associates indicates.  

The global biomanufacturing industry is growing, and the need to benchmark the relative strength of existing and emerging manufacturing clusters is becoming imperative. So as part of our 20+ years of evaluating and benchmarking the biopharmaceutical manufacturing industry (1), BioPlan Associates is sponsoring the Top 1000 Global Biopharmaceutical Manufacturing Facilities Index. We will be presenting this as an industry-driven website at the BIO International Convention in Washington DC in June (updates will also be available at 

Table 1: Top 1000 biopharmaceutical facility counts by region as of May 2011. 

Some in the industry find it hard to imagine that there are 1000 biomanufacturers worldwide. But our research, reports, and studies have shown that in China and India alone the number of qualified facilities has grown more than 20% in the past three years to their current number of 91 and 99, respectively (see Table I). To create the BioPlan BioFacility Index and rank biomanufacturers, we analyzed overall fixed capacity, manufacturing-related employment, and number of commercial and clinical products. The result is an indexed ranking based on these factors. Shortcomings in this methodology include: difficulty in establishing equivalence between capacity for, say, a monoclonal versus microbial, versus vaccine production; questions regarding which functions are "manufacturing related"; weight to be applied to each factor; and the need to monitor each factor regularly to ensure up-to-date ranking. Prior efforts to benchmark manufacturing have generally failed because managing thousands of data points, year after year, becomes virtually impossible. For this effort, we are making use of the industry's own knowledge. Our analysis uses publicly available information that we have amassed over decades. In addition, we have recruited hundreds of participants to maintain the listing. But most uniquely, to keep the data accurate, we have implemented a "Wiki" approach whereby industry is able to contribute public knowledge.  

In addition, BioPlan will pass proposed changes to the database through our review panel to ensure accuracy and consistency of any updates.  

Biopharmaceuticals represent a $135-billion industry worldwide and are growing at 20% annually, much higher than most other industries. This dynamic segment provides well over 400 drug products to the major US and European markets, and hundreds of others marketed internationally. The biopharmaceutical industry is one of the most important engines for technology growth worldwide. In fact, mainstream pharmaceutical companies have been moving into biopharmaceuticals in search of future products and profits. Currently, about 15% of worldwide pharmaceutical revenue comes from biopharmaceuticals (2), but this will grow to about 40%, with an estimated 40% of pharmaceuticals currently in development being biopharmaceuticals. The biopharmaceutical industry is maturing, and many products are coming off patent, with biosimilar versions on track to enter US, European, and lesser-regulated international markets. Thus, the number of biopharmaceutical manufacturers and facilities are expected to multiply rapidly in coming years.  

In BioPlan's Top 1000 Biopharmaceutical Facilities Index, we will provide not only counts, but also indexed rankings of each company, as well as the regional ranking based on the sum of each individual facility BioIndex. This index will provide a quantitative analysis of the strength not only of the biomanufacturers, but of the regions and clusters, as well.  

WHY RANKING AND         INDEXING IS IMPORTANT

Biopharmaceutical manufacturing facilities are strategic assets, both for the companies involved and for the cities, states, and countries where they are located. This status is unlike the financial and other industries where products and profits are often more virtual. Biopharmaceuticals involve real products, with their manufacture requiring the highest technology, trained professionals, and expensive infrastructure. Research and development (R&D), on the other hand, is increasingly diffuse, and often involves multiple facilities, outsourcing, licensing in, and collaborations. In contrast, biopharmaceutical manufacturing provides sources for revenue and profits and is physically concentrated, generally performed in a single major or, at most, several redundant facilities. Each facility typically costs more than $100 million, employs hundreds of manufacturing professionals, and provides for its community related tax and other local benefits. While biopharmaceutical R&D facilities may come and go, for example, through corporate changes, acquisitions, and bankruptcy, manufacturing facilities are physical assets that tend to continue to manufacture products despite corporate changes.  

However, technological advances are reducing bioprocessing costs and complexity, therefore lowering the bar for entry, including in lesser-developed countries. Improved expression systems and genetic engineering continue to increase productivity. Single-use bioprocessing systems are reducing the need to support infrastructure, reducing up-front investments, and are making manufacturing simpler, more affordable, and flexible.  

BioPlan is tracking more than 1000 facilities worldwide with at least some involvement in biopharmaceutical manufacture. Included are hundreds of product manufacturing facilities and many more operating at smaller scale to support R&D and clinical testing. These facilities account for approximately $10 billion in equipment and consumable sales annually. Despite its strategic importance, this information resource to track biopharmaceutical manufacturing facilities and benchmark their economic contributions is the first to provide comprehensive indexing and ranking.  

Knowing where the key people and facilities are is critical for decision-making. Besides comparisons between facilities and companies, the Top 1000 enables geographic, indexed comparisons on a local, state, or country basis. The clustering of related facilities and professionals in the same locale is important for high-tech industry growth. Companies, investors, human resources departments, analysts, regional business development authorities, and governments can use this resource to address a variety of strategic issues. For example:   

Exactly where are biopharmaceutical manufacturing capacity and employment clustered? 

Where is the industry growing the most, and by which indexed factors? 

Just how many facilities are there in China, India, Russia and other developing countries; how rapidly are these facilities growing, and how do they compare with those in the US and Europe? 

Where is large-scale capacity being added, and who controls this? 

Knowing where key biomanufacturing facilities are located around the world is essential for decision-making.

BioPharm International-06-01-2011