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Biopharmaceutical processing has been greatly improved over the past 10 years with the introduction of single-use/disposable technology. There has been a landslide of disposable products introduced in the areas of filtration, mixing, sampling, and transferring solutions aseptically. It is common to find papers and studies that detail the efficiencies that disposables offer to the biopharmaceutical market. What about aseptic filling? More importantly, what about the cost of aseptic filling? PDC aseptic Filling Systems offers an answer.
The biopharmaceutical industry faces numerous cost pressures in today’s growing market. One such pressure point is aseptic filling. The question is how to reduce cost and increase production without compromising quality. Disposables are part of the answer, but many single-use products are expensive.
Some industry visionaries have begun to explore how “lean manufacturing” principles can help improve production efficiencies and reduce cost. Lean manufacturing practice...
Upfront Chromatography A/S is a Danish company with significant expertise in the development of customized industrial processes. Successful operation of industrial separations provides proven scalability for biopharmaceutical manufacturers. Upfront Chromatography’s Rhobust™ technology has been developed to maximize the effectiveness of capture adsorption steps, using second-generation expanded-bed technology.
Rhobust™ consists of
A key advantage of the Rhobust™ platform is that it can be used directly with unclarified feedstocks. The chromatographic adsorbent captures protein molecules in one simple downstream process. Neither clarification nor adsorbent packing and repacking is required; it enables NaOH cleaning (at high temperatures) as well as enhanced selectivity upfront in the process.
Cost-Effective and Robust Processes with Rhobust™
For pilot plants and other multiproduct facilities, no presanitization, extensive prequalification testing, CIP, or cleaning validation are required. Rhobust™ Flex disp...
While industry is rapidly shifting from stainless steel vessels and hard-plumbed processes to single-use bioreactors and TFF filters, the transition between the multiple process steps remains complicated and time-consuming, often requiring fluid transfers, disrupting sterile barriers, and cleaning protocols. The simple solution to a complex process is connectivity: a continuous flow-path that interconnects all the process steps by means of a sterile singleuse process manifold. Spectrum Laboratories incorporates process filters, reservoir bags, flexible tubing, and other critical elements into a custom-designed single-use KrosFlo MBT (Module-Bag-Tubing) system to provide total product isolation in a continuous flow-path and maintain a sterile barrier through multiple process steps. Alternatively, module-tubing sets can be designed for use with bags already validated.
Custom Kros Flo® MBT™ for Serial Separation
A common complex process is serial separation in which sequential filtration steps are performed ...
Biopharmaceutical companies continue to wrestle with how to approach the implementation of process analytical technology (PAT). The accelerated adoption of many new technologies has benefited from the identification of a “killer app,” an early application that enables a readily achieved breakthrough in capability. This can be a totally new capability (e.g., e-mail on the internet) or a major step-change (e.g., 10× or more) in an existing capability. Flashes of inspiration cannot be achieved at will, but we can identify elements of existing processes that are ripe for such breakthroughs. For example, downstream processing capacity increases have fallen far short of the 10× − 1,000×improvements recently achieved in upstream titers due to the inability to increase buffer capacity within existing floor space. The current paradigm for buffer preparation is to premix and store buffers in (relatively large) tanks. A new paradigm is required.
Two candidate solutions are proposed:
The first solution implies moving...
Figure 1.
At recent scientific conferences on downstream processing, there was much discussion about the need to reduce process volumes. Many manufacturing sites are space-limited and to take advantage of chromatographic process improvements, they must be made without the need to increase buffer and holding tankage.
One significant opportunity to increase process throughput is to replace conventional with higher capacity ion exchange resins. Recently developed high capacity resins allow more protein to be loaded without the need to replace existing column hardware for larger column dimensions. To keep pace with increasing titers of feedstock, using higher capacity resins results in process improvements while utilizing existing chromatography hardware.
This article demonstrates that an even better solution is available in the form of new Toyopearl GigaCap resins (specifically designed for packed bed use) which have both high capacity and improved elution kinetics. When combined together and depending on yo...
Bioengineering AG has 30 years of experience in the construction of hygienic plants. Thereby holistic process equipment from the feed line to the valve system for the final cleaning has become the status quo.
Our new BioAccessories department focuses on all our components and accessories. Now you do not need a Bioengineering system to be able to use our developments. For special applications we offer the possibility of custom-made or even newly developed parts.
Safety with Leak-Proof Couplings:
Sterility begins in the pipelines, and flexible couplings are often a source of contamination. For maintenance, several couplings are necessary in each plant. Sterile couplings from Bioengineering provide safety without deadlegs and crevices and with seals flush with the pipe wall. Our diaphragm valves can be fitted with sterile couplings as well.
One-Way and Multiway Valves for Every Application:
For over 10 years Bioengineering AG has developed and produced diaphragm valves of all sizes with various actuators a...
Figure 1.
The Newmix-Levtech Magnetic Mixer system is a compact and noninvasive single-use mixing system. Its heart is a mixing bag that incorporates an innovative, bottom-mounted, magnetically driven impeller capable of providing efficient high-torque mixing for all powder-liquid and liquid-liquid mixing applications. The impeller rides on a low-friction, inert bearing assembly designed to ensure low particle shedding while allowing mixing of very high powder loads in large liquid volumes. All product-contacting surfaces are 100% disposable.
Introduction
Powder-liquid mixing is a common requirement in biopharmaceutical processing. To maximize mixing efficiency for powder-liquid applications, the Magnetic Mixer is available with a large (6.35-in.), centrally located magnetic impeller. In this experiment, a Magnetic Mixer system’s ability to suspend a very high powder load and its resistance to stalling when buried by compacted solid were tested. The powder chosen was diatomaceous earth — a fine, inert, an...
The increasing popularity of “single-use” or disposable bioreactors for upstream processing can be understood by considering a typical biotechnology manufacturing facility. The infrastructure required to implement traditional glass/steel bioreactors is substantial, and the time and expense required to validate it can be significant. The requirement that both the bioreactor itself and its ingress and egress tubing use inert materials such as 316-L electropolished stainless steel results in a large capital investment and long lead times for components. In addition to mitigating the capital cost of steel systems, single-use systems bring the additional benefits of shorter preparation and sterilization times, lower purified water volumes for cleaning vessels between growth runs, and significantly reduced maintenance times. Additionally, single-use bioreactors and the associated plastic tubing lend themselves to being reconfigured and validated quickly and efficiently as manufacturing or process requirements c...
Figure 1.
Select promising leads earlier in the development process with the Octet RED system’s rapid, highly sensitive detection and label-free analysis of small peptides and small molecules. The Octet RED system’s rapid data acquisition and high sensitivity means that even the fastest reactions and smallest binders will be easily characterized.
The Octet RED system shares the advantages of the Octet QK system: BLI technology, no microfluidics, eight-channel parallel processing, walk-away automation of up to 96 samples, and affordability — while offering expanded detection. By monitoring eight independent interactions simultaneously, the system also increases productivity without additional resources or manpower. The system’s reasonable cost keeps assays in-house with fast, accurate results.
Principle
Protein binding and dissociation events can be monitored by measuring the binding of one protein in solution to a second immobilized on the ForteBio biosensor.
Multiple Analysis Modes
Biosensor Surface Chem...
Throughout its 50-year history, WL Gore & Associates has earned its reputation for providing innovative, high-value products such as GORE-TEX® outerwear, Elixir® guitar strings, and GORE-TEX® vascular grafts. Having invented expanded polytetrafluoroethylene (ePTFE), Gore offers an unparalleled level of technical knowledge and experience with this versatile material. This know-how starts with a deep scientific understanding of fluoropolymers, particularly PTFE processing and fluoropolymer development. Gore’s unique technical agility enables the development of a wide range of processes, products, and creative, reliable technologies.
Performance Characteristics
Gore’s configuration engineering of ePTFE and other materials can result in products that offer a wide range of performance characteristics, including biocompatibility, chemical and thermal resilience, low dielectric constant, antibacterial functionality, odor absorbency, radiation resistance, selective permeability, transparency/opacity, dimensional ...
Figure 1.
As the pharmaceutical industry focuses on developing biotechnology-derived drugs, bioanalytical groups must produce generic methodologies for antibody characterization with higher throughput and faster sample turnaround times. Although antibody selectivity varies, overall antibody structures are highly conserved. Intact antibody LC/MS analysis is useful for profiling glycovariants and C-terminal Lys processing, and even more information can be gleaned from analysis of a reduced antibody.
UltraPerformance LC® (UPLC®) technology permits efficient desalting and rapid LC/MS mass profiling of an intact (4 min) and reduced (10 min) murine IgG1 monoclonal antibody. Highresolution mass spectrometry provides a powerful approach for profiling batch-to-batch variations and studying structural changes associated with drug production, formulation, and storage. These methodologies yield a simple, holistic view of the molecule and its variants by providing intact mass information that can be used to confirm pr...
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The pharmaceutical industry has relied on laboratory testing for all of the past century. Typically, biopharmaceuticals are manufactured in batches and tested for quality using off-line laboratory test methods of samples extracted from the process line.
However, taking the laboratory to the sample changes the way analyses are conducted. The FDA Guidance for industry,
PAT: A Framework for Innovative Pharmaceutical Development, Manufacturing, and Quality Assurance
, promulgated in final form in September 2004, encourages the use of available process analytical tools to support innovation and efficiency in biopharmaceutical processing. The intent of the guidance is to describe a scientific, risk-based regulatory framework to support innovation and efficiency in biopharmaceutical development, manufacturing, and quality assurance. The PAT framework is a system for designing, analyzing, and controlling biopharmaceutical manufacturing through timely measurements of critical quality and performance att...
Microfluidics offers its M-110P Microfluidizer® processor, a benchtop laboratory machine that requires no compressed air or cooling water for hydraulics. Designed to operate reliably, quietly, and efficiently at up to 30,000 psi process pressures, the portable M-110P requires only a standard 20-amp, single-phase “household” electrical outlet and can easily be incorporated into any laboratory setting.
Using Microfluidics’ fixed geometry diamond interaction chamber technology and ceramic plunger, the M-110P easily enables the production of nanosuspensions and nanoemulsions, as well as liposomal encapsulation and cell disruption with the fewest number of passes. Additionally, the M-110P allows flow rates between 110 mL/min and 155 mL/min depending on process pressure—which are higher flow rates than any comparable, competitive product. These flow rates enable the efficient processing of pharmaceutical formulations, fine chemicals, and biological material ranging from simple oil-in-water emulsions to highly i...
Who Is Advanced Scientifics?
Advanced Scientifics is an ISO 13485:2003, FDA registered global leader in the design and manufacture of single use systems for the Pharmaceutical and Biotech industries. Our vast array of manufacturing capabilities allows us to provide a custom disposable system in some of the industry’s shortest lead times. From system design, to the highest standard in quality production and assembly, to final packaging and sterilization, advanced Scientifics is your complete single use systems provider.
Facilities
Advanced Scientifics holds two manufacturing facilities, headquarters and main manufacturing located in Millersburg, Pennsylvania as well as off-shore manufacturing capabilities located in Matamoros, Mexico.
The Mexico facility duplicates the US facility’s capabilities including high volume manufacturing, class 10,000 clean rooms, as well as the same strong quality system, sterilization capability, and is ISO 13485:2003 certified and FDA registered.
Single Use Systems
Advanced Sc...
Applikon Biotechnology BV (in the Netherlands) and Aber Instruments Ltd. (in the United Kingdom) have entered into an agreement for the exclusive supply of the first integrated bioreactor controller and sensor for viable cells on the market. Applikon Biotechnology will offer the viable cell sensor and controller option in its EZ Control bioprocess controller portfolio designed for the biotechnological and biopharmaceutical industries.
The Aber Biomass Monitor uses a unique
in situ
probe that incorporates four annular ring electrodes to apply a radio frequency field to the biomass. Electronic processing of the resulting signal produces an output which is an accurate measurement of the concentration of viable cells. The system is responsive to cells with intact cell membranes and is insensitive to gas bubbles, cell debris and cells with leaky membranes.
Aber Instruments, celebrating its 20th anniversary in 2008, has upheld its tradition of innovation and staying ahead of the competition by supplying clien...
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AmProtein Corporation is a biotechnology powerhouse that has revolutionized the biologics industry with the discovery of a novel expression vector and development of a disposable bioreactor series. AmProtein has used these two discoveries to develop an unbelievably simple and efficient, industrialized CHO cell protein production platform. This platform includes a revolutionary expression vector; a single-use, plastic bag-based bioreactor system based on a novel, patented O
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transfer method; and serum-free cell culture media. The platform is scalable from bench to manufacturing, and our bioreactor system makes current bioreactors utterly obsolete. AmProtein currently uses this platform in its contract manufacturing business, which is especially suited for production of difficult-to-express proteins. AmProtein is successfully applying its vector and manufacturing platform to plant, insect, yeast, chicken, and fish cells.
AmProtein is open to fostering partnerships around the globe for the codeve...
Figure 1.
The costs of producing monoclonal antibodies (MAbs) from mammalian cells are extremely high. Producing 500 kg/year of a MAb can run $200,000,000/year or more. Great effort has been made by the biopharmaceutical industry to reduce these costs. Companies have improved or expanded their existing infrastructures for MAb manufacturing, optimized media formulations and feeds, and/or genetically engineered production cell lines, with the current industry goal of 10 g/L titers and productivities of 100 picograms/cell/day. A twofold increase in volumetric productivity can easily be expected to provide savings of 20% or more in costs of goods sold. Increases of this magnitude translate to costs savings in upstream biopharmaceutical production amounting to 6–10% of total sales.
The CDI Bioscience (CDI) solution to improving upstream yields is a patented and innovative technology to increase the yield of bioproducts manufactured in cellular systems. Genetic engineering is used to introduce factors that, whe...
Biopharmaceutical manufacturers face increased pressure to commercialize new drugs faster and at a lower cost. As a result, production demand is outpacing available manufacturing capacity. To meet existing and future demand and maintain a competitive advantage, many manufacturers are striving to develop increased efficiencies within their manufacturing processes. This is where single-use systems — particularly single-use transfer lines — can deliver significant value, including added flexibility, improved production yields, and increased cost savings.
The trend today is toward transfer lines or tube sets that use single-use connectors and filters to achieve sterile media transfer between process equipment or between production suites. A key benefit of single-use transfer lines is their ability to help boost productivity and accelerate time to market by reducing the downtime associated with cleaning and validation. This, in turn, helps reduce operational expenses by minimizing labor, chemical, water and en...
Emerging lifestyle diseases and an aging population are creating a number of major health challenges. Over the past few years, the food industry has been more focused on functional foods, those intended to be consumed within the normal diet but containing biologically active components with physiological benefits intended to reduce certain chronic diseases. This US$35 billion market has been steadily growing at 8-14% per year, as estimated by the Food and Agriculture Organization of the United States (FAO). Although the United States and Japan hold the largest markets in this industry, China is expected to double its share to US$12 billion/year by 2010.
Functional Food the: New Sphere of Action
Recent research shows that diet can affect the formation and course of diseases such as cancer, cardiovascular disease (CVD), osteoporosis, and Alzheimer’s disease. As a result, additives such as vitamins, fatty acids, probiotics, bioactive peptides, and antioxidants are added to food products to increase their fun...
Figure 1.
The demand for biopharmaceutical products is increasing rapidly; however, development timelines of about 12 years leave little time for return on investment before a patent expires. Fast process development is therefore crucial to speed up the market entrance of new biopharmaceuticals.
Until recently, chromatographic process development was restricted to a sequential regime. However, new innovative methods have been developed to perform up to 96 parallel chromatographic experiments on a Tecan Freedom EVO® robotic workstation (pictured, right), allowing for thermodynamic, kinetic, dynamic binding, and elution studies in an automated, parallelized, and miniaturized mode, which saves time, money, and target molecule.
Here we present an overview of the most important chromatographic process development tools on Tecan Freedom EVO® robotic workstations: finite bath thermodynamic studies (
1
), batch binding in PreDictor™ plates from GE Healthcare, bind/elution studies, and breakthrough/elution experim...
Figure 1.
Manufacture of synthetic peptides ranges from milligram to multikilogram amounts. When a peptide is a biopharmaceutical (API) candidate, the amount required will increase as it moves through clinical trials to marketed product. When developing purification methods, the required “product” API quantity and purity should be considered.
Bradykinin is a physiologically and pharmacologically active peptide of the kinin family used in development of antagonists and therapies for hereditary angioedema. Here we describe two different methods of purifying a crude sample of synthetic bradykinin on a preparative scale.
Experimental
Following analytical method development on a 250 mm × 4.6 mm PLRP-S 10-µm 100Å column, isocratic separation conditions of 0.1% TFA in 21:79 ACN/water were chosen at a flow rate of 360 cm/hr. For a 50-mg purification of crude bradykinin, a 1-in. dynamic axial compression (DAC) column was used because it scales to full production. Therefore, a 250 mm × 1-in. Load&Lock™ (L&L) column...
+1 Improving production quality, efficiency, and yield is the objective of every commercial biotechnology process. Automation of assay tasks now routinely performed manually directly improves production bioprocesses. The US FDA supports and encourages this activity through the process analytical technology (PAT) initiative within its regulatory structure. The features and benefits of the online series of Automated Reactor Sampling (ARS-M™) systems supplied by Groton Biosystems are detailed here.
Online Automatic Assays
Today the most vital late-stage and production assay measurements — with the exception of basic temperature, dissolved gasses, and pH — continue to be performed using off-line, manually operated laboratory instruments. For example, amino acid analysis, cell viability, and nutrient monitoring are typically performed using traditional off-line instruments. This leads to inevitable delays and creates the potential for errors in sample handling. The historical nature of these data would be difficu...
Figure 1.
Single-use components are finding more use in the biopharmaceutical industry. One application increasing in use is transportation of sterile liquids. There are many challenges to consider when shipping a flexible container with sterile liquids. Typical Bio-Containers used in shipping applications are constructed of multiple layers. The fluid-contact layer is typically constructed of low, linear low, or ultralow density polyethylene (LDPE, LLDPE, or ULDPE). A variety of gasbarrier layers can be used in construction of the multilayer film, and one typical is ethylene vinyl alcohol (EVOH). The outer layer provides strength and can be constructed of polyethylenes, nylons, or polyesters.
Although the film construction is complex and engineered specifically to endure the harsh conditions it may encounter, Bio-Containers manufactured using plastic have property challenges, such as low penetration resistance, when compared with stainless-steel vessels. These containers must be protected during movement ...
Figure 1.
There are a variety of affinity tag-protein strategies, and the use of affinity tags depends on the target protein, the expression system, and the application. The Octet QK and Octet RED systems provide an easy-to-use, label-free, high-throughput system to capture affinity-tagged biologics for quantitative or kinetic biomolecular interaction analysis. Kinetic characterization of biologics during clone selection, screening, and development is critical at all stages of therapeutic engineering and bioprocessing.
Key Applications
Principle
Protein binding and dissociation events can be monitored by measuring the binding of one protein in solution to a second protein immobilized on the FortéBio biosensor.
Key Features
Key features of the Octet QK and Octet RED systems for kinetic screening and characterization include the following:
Figure 1.
Developed in 1996, the WAVE Bioreactor™ and Cellbag™ disposable bioreactor systems have revolutionized cell culture for commercial and R&D applications in the production of cell-derived products using animal, insect, plant, microbial, and virus cultures. Oxygen transfer and mixing are accomplished by the novel principle of wave-induced agitation. Wave action generates free surfaces for bubble-free oxygen transfer from the headspace of each Cellbag bioreactor, and it also mixes the fluid suspending cells or particles. This design eliminates the need for mechanical mixing or gas sparging, both of which introduce shear damage to cultures. Air is continuously purged through the bioreactor headspace to supply oxygen and remove metabolic gases. This system can be adapted to control pH and dissolved oxygen or operation of perfusion cultures.
Optimizing Immunoglobulin Expression in a WAVE Bioreactor
Joseph Mollick, MD, PhD, of the Stanford School of Medicine’s Division of Oncology studies tumor-specific...
Figure 1.
On-line biomass monitoring using capacitance is now a routine task in cell culture. The capability to develop advanced bioprocesses relies on sensitive process monitoring technologies and robust process control strategies.
Unlike optical technologies, capacitance is insensitive to dead cells, cells debris, gas bubbles, or other matter in suspension. With its capacitance-based sensor, FOGALE nanotech has supplied many pharma/biotech companies around the globe. The Biomass system is widely recognized as a powerful tool from process development to CGMP production scale. It is ideal for monitoring animal cells in suspension (including clumping cell lines) or attached to microcarriers and is also routinely used for yeast or bacteria batch and fed-batch processes, with high biomass concentration up to 200 g/L dry weight.
Figure 1 demonstrates how the Biomass system is reliable for on-line tracking of viable biovolume by measuring capacitance in a fed-batch CHO culture.
Capacitance measurements can als...
Cell counting and viability assessment are routine but essential parts of bioprocessing procedures, and accuracy, reproducibility, and standardization are essential to successful outcomes. Traditional methods produce variable results because they rely on subjective discrimination of dye-stained cells using a hemacytometer and microscope. Substantial operator-to-operator variability in the cell counts is common, even when one researcher examines the same sample twice.
Guava technologies offers a solution: a simple, accurate, and reproducible absolute cell counting and viability assay that overcomes many of the drawbacks of the manual method. The simple and robust Guava ViaCount® assay eliminates statistical variation by sampling large numbers of single cells, and it removes operator variability by categorizing cells as live, dead, or apoptotic by quantifying fluorescence staining.
Accurate Cell Counting via Microcytometry
The Guava ViaCount assay runs on all of the compact, benchtop Guava Systems: PCA™, PC...
innovatis has established a technology portfolio to assist with the implementation of process analytical technology (PAT) at pharmaceutical companies focused on manufacturing biologics. PAT is a critical component in FDA’s initiative entitled, “Pharmaceutical CGMPs for the 21st Century: A Risk-Based approach,” which was announced in August 2002. This initiative is intended to support innovation and efficiency in pharmaceutical development, manufacturing and quality assurance. PAT is a system of building into processes “quality by design” (QbD), an idea that product quality begins with a deep and sophisticated understanding of the scientific and engineering principles involved in a manufacturing process and the identification of variables that affect quality. Product quality cannot be “tested into” a product; it must be built in from the beginning.
Our goal is consistent with QbD and PAT: innovatis wants to help modernize the way laboratories use and manage their sample monitoring data. Because monitoring ...
Invitrogen has 45 years of experience in the cell culture business. Over this time, the Invitrogen name and reputation has made us the recognized leader in cell culture systems. In 2003, Invitrogen expanded on this tradition in innovative cell culture products through the creation of a team focused on process development, PD-Direct® BioProcess services. The PD-Direct team responds to core customer needs by offering customers comprehensive service from cell line and media development, to process optimization.
Upstream Optimization Services
The PD-Direct team brings together the best scientists and technologies in upstream optimization. A proven approach of applying high-throughput technologies to process design within an integrated workflow around media and feed optimization yields performance improvements in your media. Our ability to bring this step change in performance comes from several different elements:
Cell Line Services
The science of cell line development has become complex. Increasing productio...
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Established in 1997, integrated Genomics (IG) provides a full range of products and services to support research in microbial genomics, metabolic analysis, and engineering. Our flagship genome analysis system, ERGO™, provides a bioinformatics toolkit enabling better and faster identification of gene function and pathways across organisms, which is evident within more than 100 scientific publications in top peer-reviewed journals (www.integratedgenomics.com/igpubs.html).
The ERGO in silico metabolic discovery and analysis tool set is the world’s largest commercially available database of microbial genomes coupled with a state-of-the-art analytical tool set. More than 900 bacterial genomes have been curated manually and associated with a network of biochemical pathways. Currently, ERGO contains more than 3.6 million open reading frames, of which 2.5 million have functional assignments and 6,500 metabolic pathways grouped into subsystems, all linked together with data mining and analysis tools. ERG...
