Much has already been written lately about addressing the so-called “downstream bottleneck(s).” A number of companies are leading the way toward developing products and platforms for reducing both the costs and the time required for downstream processing. Our task with this special issue was to provide a state-of-the-art update on these activities — but as always, within a limited number of pages allotted.
The primary issue behind this bottleneck debacle is to address purification challenges posed by aggregation in cell culture supernatants after cells are pushed to high antibody titers. This special issue therefore touches upon trends in single-use purification options, single-stage and turn-key harvest solutions, and cutting-edge innovations in chromatography.
Knowing that our coverage would only scratch the surface of the work being done, I polled members of BPI’s editorial advisory board. I asked them what sorts of articles they want to see more of related to current trends in downstream processing. T...
Humor columnist Dave Barry once opined, “Without question, the greatest invention in the history of mankind is beer. Oh, I grant you that the wheel was also a fine invention, but the wheel does not go nearly as well with pizza.” Certainly, few biological concoctions have achieved that level of popularity — but with a little luck, your company will produce at least one invention with the potential for measurable success in its target market. But with success comes competition, and that’s where protecting your invention becomes important.
We’re living in a time when donating intellectual property to the pubic is trendy: Linux,
Wikipedia,
and Creative Commons are just a few examples. For many readers of
BioProcess International
who have dedicated their careers to improving the human condition, altruistic concepts such as these may be inherently appealing. However, in the absence of sufficient public funding or philanthropy, the high cost of developing effective biopharmaceuticals must be funded by sellin...
What’s keeping senior biopharmaceutical executives awake late at night? According to BioPlan Associates, Inc., which publishes an annual comprehensive survey of the state of worldwide biopharmaceutical manufacturing, capacity constraints are among the key issues at hand (
1
). And one of the most important constraints is the lack of physical capacity in purification equipment. Bioreactors are producing a lot more protein than current downstream purification steps are designed for. Overcoming the resulting bottlenecks may require increasing the productivity of downstream unit operations and turning over processing equipment faster to handle each consecutive batch.
Many biopharmaceutical manufacturers are turning to single-use systems to increase product throughput. Recent advances extend the use of disposables from the typical bioreactors and buffer or media storage bags to clarification and chromatography steps – and even to viral filtration and drug formulation. With such advances, the industry is on the...
As organizations launch operational excellence (OpEx) programs, they are faced with challenges that must be overcome before they can achieve true excellence. One of the largest barriers to overcome is employee perception. The first step is to provide training that will energize employees and change their understanding of what operational excellence can do for them and their organization. For this reason, Bayer has developed an OpEx fundamentals training that starts with the “heart and mind,” focusing on less-threatening “soft skills” first and thus opening an audience to listening to the message the team is trying to convey.
Our training also focuses on neurolinguistic programming (NLP) and Lewin’s change model. This helps remove misperception and retrains thought development so employees can eliminate false perceptions and replace them with new understanding and openness to operational excellence. Although team members are not instantly converted, the process of change is triggered so that continuous imp...
BioProcess International
has followed, from the beginning, the ways in which single-use technologies have transformed the landscape of industrial bioprocessing. On 18 March 2009, we organized a panel session at the annual Interphex conference (Jacob Javitz Center, NYC) to drive discussion toward longer-term implications of single-use components and technologies on the future of bioprocessing. Is their use a cost-saving strategy overall? What economic factors are driving their adoption? The panelists were prepared to address such topics as economic considerations in early planning stages, risk management and supply chain issues, the relationships between suppliers and users, factors that determine which processes are good candidates for disposable products, and some implications of wider adoption of single-use technology when considering total life-cycle costing. My comoderator Jerry Martin (see the Panelists box on the next page for participant details) and I each directed questions to the panelists. The...
Manufacturing processes for biopharmaceuticals have undergone significant changes over the past decade. One of the most striking results of improved process sciences is the dramatic rise in expression levels from animal cell cultures. Figure 1 shows how some monoclonal antibody titers have increased about 30-fold over the past 15 years. These increasing titers have allowed current biomanufacturing facilities to produce larger product quantities than anticipated at the time they were designed and built.
Figure 1:
As a result of those increasing titers, the “bottleneck” in biopharmaceutical production has shifted from upstream production processes toward downstream processes (
1
). In particular, chromatographic capture is currently presenting significant challenges in terms of facility throughput. The mass of product that needs to be captured during each batch has often exceeded the maximum binding capacity of a single column, so capture columns generally are being cycled multiple times during a single bat...
+2Fast, precise, and accurate quantification technologies are indispensable for efficient process development in applications such as IgG production in a GXP environment. Based on surface plasmon resonance (SPR) technology, the Biacore C system from GE Healthcare (
www.biacore.com
) is an alternative technology for IgG quantification that has benefits over traditional methods. Assay development is simplified and accelerated due to real-time detection. Assay hands-on time is reduced, and sample throughput can be increased using automation and efficient data evaluation with regulatory-compliant software. We developed a generic platform for hIgG quantification using the Biacore C instrument.
Materials And Methods
Immobilization of Mouse Anti-hIgG:
Mouse anti-hIgG (antihuman Fc) was covalently bound as a ligand by means of primary amino groups on the carboxy-methylated dextran molecules of a sensorchip surface. We performed all our measurements using research-grade Biacore CM5 chips. Mouse anti-hIgG was immobi...
+1 Over the past 10 years, disposable bioreactors have grown from a niche tool servicing small-scale projects to a common and essential component in the CGMP production of human therapeutics (
1
). Recent advances in filter integration, aseptic connectors, and disposable sensing allow entire cell culture processes to be performed using only single-use components. However, harvest and clarification operations remain largely dependent on centrifugation, cross-flow filtration, and depth filtration (
2
), which are all techniques that have not been widely adapted to single-use implementation. Their use in harvesting from disposable bioreactors can lead to process bottlenecks (
3
), especially when large numbers of relatively small bioreactors are in use (e.g., for clinical and research laboratories). In such cases, the advantages of ease-of-use, disposability, and turn-key processing often outweigh the need for optimized, molecule-specific processes.
Here I report the results of a three-phase, nine-month–long st...
We have previously described a patented nonsparging, nonbubbling oxygen transfer method (
1
). This method is based on interaction between the air-exposed smooth surface of a bioreactor vessel and culture medium repeatedly sweeping across it with a certain force, which seems to generate microscopic bubbles among the water molecules (
2
). We manufactured high–oxygen-transfer Current suspension bioreactors with working volumes of 5 L, 50 L, 150 L, and 300 L.
Here we describe the use of these suspension bioreactors as “artificial lungs” or dissolved oxygen generators to seed and irrigate cell columns that are completely filled with affordable, nonwoven polymer fibers (made in China by a member of the AmProtein alliance) to achieve high-density culture. These bioreactors thus mimic human heart and lung machines to perfuse a cell column that is completely filled with polymer fiber discs used as cell anchorage carriers. The Current bioreactors provided for high-density cell culture of various cell types, inclu...
+17 Biopharmaceutical manufacturing is divided into two areas: upstream fermentation or cell culture and downstream purification processes. Each area contains multiple unit operations. A unit operation is defined as a step in processing using a particular type of equipment. Here, we focus on downstream process development, which must reliably produce a highly purified drug substance (often >99%). Downstream processing includes recovery, capturing, and polishing steps.
The primary downstream unit operation is chromatography because of its simplicity and high resolving power (
1
). Most process trains involve at least two distinct, orthogonal chromatography steps chosen from the number of chromatography supports available with various ligands attached to solid matrix. Affinity, cation-exchange (CEX), anion-exchange (AEX), ceramic hydroxyapatite (CHT), and hydrophobic-interaction chromatography (HIC) are the main types of chromatography modes used in large-scale bioseparations. The mode used for polishing depend...
+4 Use of automated liquid handling equipment for rapid testing and reproducible screening of thousands of molecules, cells, and compounds has become an essential component of life-science laboratories across the globe. Along with an increase in such use, transferred volumes have shrunk, as demands increase on transfer accuracy and precision when aspirating, diluting, dispensing, mixing, and washing. Automated liquid handlers are generally used to increase the productivity and repeatability of volume transfer, but as discussed here, they are still prone to error. So it is important to understand how some errors can be recognized and prevented to maintain liquid handling quality assurance, especially when transferring critical reagents.
Because concentrations of biological and chemical species are volume dependent, the accuracy and precision of individual (stepwise) volume transfers directly affect the amount of critical reagent transferred to or from an assay. Inaccurate or imprecise delivery can easily resu...
For many years, biopharmaceutical manufacturers have worked to increase capacity, address upstream production issues, and improve product yields. Notable successes recently achieved in upstream technology have significantly increased expression rates and therefore, upstream production capacities. Successes in generating higher titers combined with increasingly stringent quality and regulatory requirements have led to a number of challenges in aligning the efficiency of downstream processing with upstream titers. It is generally recognized that downstream processing costs account for about 70% of the total biomanufacturing cost (
1
). As a result, improvements in product recovery and purification are urgently needed. In trying to overcome this challenge, companies analyze where the best solutions lie: in streamlining operations, exploring emerging technologies, and/or using disposables.
Biomanufacturers are showing increasing interest and seeing strong development in single-use technologies and devices bot...
+1 Ongoing requirements for additional insight and CGMP-compliant measurement have led to interest in new technologies that can be applied to the analysis of many different types of particle-containing fluids. Micro-Flow imaging (MFI) is a robust, versatile, and intelligent vision technology that is increasingly used for evaluating populations of microparticles encountered during bioformulation development (
1
). It is also being accepted in fill–finish processes and many other biotechnology applications.
Features of MFI technology that have contributed to its rising popularity include the direct insight provided by imaging, a superior sensitivity for near-transparent particles, ease of use, and flexibility in resolving and independently analyzing different particle types present in heterogeneous populations. For example, the technology is used to quantify transparent protein aggregates of various morphologies that can coexist with silicone droplets, air bubbles, micelles, and a variety of foreign contaminan...
+1 IBC’s BioProcess International Conference and Exhibition (BPI) will head back to the US east coast for 2009 to Raleigh, NC. This has become the largest, most respected forum focusing on technical, regulatory, and strategic planning challenges in the manufacture of biotherapeutics. Four comprehensive conference tracks as well as intimate workshops, strategic discussion groups, training seminars, and colocated conferences will continue to provide you with an interactive and lively format all in one location. BPI allows you to meet face-to-face with industry experts and colleagues across various disciplines of biopharmaceutical manufacturing — to get the answers you need to move your company forward. An expanded exhibit hall (which was sold out in 2008) will showcase the latest array of technologies and services from more than 150 exhibitors, giving you the chance to compare and evaluate the best products on the market.
In addition to the regular conference programming, all BPI Conference attendees will have...
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In November 2008 the US FDA finally issued a new draft guidance on process validation (
1
). The original guidance on this topic was published in May 1987, and the FDA explained that “since then, we have obtained additional experience through our regulatory oversight that allows us to update our recommendations to industry on this topic.” The new guidance is intended to reflect some goals of the FDA’s Pharmaceutical GMPs for the 21st Century, an initiative that was finalized in 2004. This update has been long anticipated.
The new draft is to be applied to manufacturing human and veterinary drugs, biological and biotechnology products, finished products and APIs. Its key principle is alignment of process validation activities with the product development life-cycle and the use of quality by design (QbD). Collection of scientific data and demonstrating process capability by statistical analysis are emphasized.
The draft guidance proposes splitting validation into three stages: process design (based on knowl...
