October 1, 2006 (Vol. 26, No. 17)
Capacity, Compliance, and Quality Remain the Keystones for Success in Biomanufacturing Operations
Laureate Pharma (www.laureatepharma.com)recently signed an agreement with LPath (www.lpath.com) to manufacture Sphingomab™, Lpath’s lipid-specific humanized Mab.
Laureate will provide the full-scale development of this product for clinical trials, including cell-line development, optimization, upstream and downstream development, and final cGMP manufacture.
“We strive to combine biomanufacturing expertise with flexibility and new technologies and capabilities,” says Michiel E. Ultee, Ph.D., senior director of biopharmaceutical operations at Laureate. “We are increasing the use of disposables, single-use technologies, higher productivity cell lines, and higher efficiency chromatography resins for downstream processes to provide efficient production from a vial of cells to vials of product.”
Expectations regarding improved manufacturing efficiencies at contract manufacturing sites worldwide were expressed by most contractors interviewed for HighTech Business Decisions’ (www.hightechdecisions.com) new report, “Biopharmaceutical Contract Manufacturing: Improved Processes and New Capacity for Pipeline to Commercial Production”.
Overall, 33 biopharmaceutical contract manufacturers reported that the number- one trend in the industry is the emergence of new technologies to increase biomanufacturing efficiencies: disposable single-use reactors, better expression systems and yields, cell-line optimization, improved media, improved drug potency, miniaturization, perfusion, improved purification systems, and better process technologies both upstream and downstream. Although the development of ever-increasing biomanufacturing efficiencies is important to contractors and their clients, capacity, compliance, and quality remain the keystones of the industry.
To meet various needs, some contractors have developed expertise in several areas to ensure an efficient pathway to cGMP manufacturing and clinical materials for each individual client. “We work with three types of bioreactors (stirred-tank, hollow-fiber, single-use disposable systems), three types of viral-removal technologies in downstream processes (dead-end, tangential-flow, and hollow-fiber), and provide aseptic filling services for fill volumes ranging from 0.1 to 100 mL for aqueous solutions, suspensions, and even some nonaqueous solutions,” Dr. Ultee explains.
The company has installed 500-L working volume disposable bag bioreactors to streamline cell scale-up into the stirred-tank bioreactors and simplify processing and turn-around time while reducing the need for water for injection. Laureate has a high-speed aseptic filling line with disposable product-contact parts, which allows for a quick changeover and fill, up to 20,000 vials. The company has also expanded its hollow-fiber perfusion bioreactors to boost production per unit by 40% while maintaining the same floor footprint.
Laureate recently announced a number of new projects, including full-scale antibody tech transfer/development and manufacturing for Seattle Genetics (www.seattlegenetics.com), Bradmer Pharmaceuticals (www.bradmerpharma.com), and LPath (www.lpath.com), and recombinant protein projects for Enobia Pharma (www.enobia.com) and Iconic Therapeutics (www.iconictherapeutics.com). To meet clients’ requirements, Laureate is growing its workforce by 15% and investing $9 million to expand its facility by year end.
Multiple Expression Systems
Diosynth Biotechnology (www.diosynthbiotechnology.com) signed a long-term commercial supply agreement with Dendreon (www.dendreon.com) to produce a bulk ancillary component for its Provenge® product. Diosynth’s experience with the baculovirus expression vector system (BEVS) production technology played a role in that agreement. Currently, the company is manufacturing in the insect cell system at the 2,000-L scale.
According to Frank Tielens, president of Diosynth Biotechnology, “We have been rewarded with projects that take advantage of our expertise in the newer systems such as baculovirus-based and Pichia pastoris expression.”
George Koch, Ph.D., CSO at Diosynth, adds, “It is likely that our ability to scale-up, BEVS in particular, as well as our international capabilities and infrastructure, influenced Dendreon’s decision to go with Diosynth.”
In addition, the company has a history working with expression systems that are standard in the industry, such as E. coli and mammalian cell culture. Diosynth manufactures both clinical material and commercially approved products. “We are manufacturing difficult-to-produce macromolecules in evolving expression systems as well as the current state-of-the-art systems, whether the drugs are therapeutic enzymes, Mabs, or other recombinant proteins,” Tielens adds.
The pharmaceutical industry has added regulatory constraints, so contract manufacturers must be extremely cautious regarding the use of new technologies. “Within these constraints, technology advances that enhance processing capabilities are used for the benefit of customer projects,” Dr. Koch says.
Diosynth increased the cooling capacity in its production fermenters and development units for optimized production processes. The company is using disposable reactors in cGMP production as part of a seed train toward the larger-scale bioreactors to save time and costs with regards to sterilization, cleaning, and validation.
The company has experience with fed-batch processes as well as perfusion processes. “Perfusion is well-suited for the production of complex glycoproteins requiring a low residence time in the bioreactor,” Jan Visser, director of small-scale cell culture and development at Diosynth, states.
A perfusion mode of operation is characterized by the continuous addition of fresh medium and the withdrawal of an equal volume of used medium while retaining the cells in the bioreactor. “We currently have several cell culture perfusion processes under development in addition to commercially producing Organon’s (www.organon.com) Puregon® perfusion-based product,” says Visser.
At Boehringer Ingelheim (BI; www.boehringeringelheim.com), high-throughput screening of microbial hosts, high-yield expression systems, and alternatives to chromatography are three strategic areas for new technologies. Roman Necina, Ph.D., head of biopharmaceutical production, says, “Efficient technology and quality are key to the success of the project, so our approach is to develop competitive technology rather than building capacity.”
The quick identification of an optimum expression system and the best-suited host or plasmid combination, in terms of quality and yield, are critical for efficient biomanufacturing. BI Austria established a high-throughput-screening approach that allows the screening of a high number of bacterial and yeast hosts in parallel. Both qualitative and quantitative parameters are evaluated.
“The expression systems we evaluate include E. coli, based on periplasmic and cytosolic production (preferred as insoluble inclusion bodies), and product-secreting yeast systems,” Dr. Necina says. “After a screening round, the host with the highest productivity and best quality is selected.”
Increasing the volumetric yield of microbial expression systems is required to achieve low cost-of-goods-sold figures. “Our new Autoprotease Platform technology based on E. coli provides high-yield expression of finicky and large proteins,” Dr. Necina explains. “The fusion partner tends to aggregate and guides the fusion partner into inclusion bodies, providing yields up to 15 g/L. Subsequent refolding is mediated by the fusion partner, which cleaves itself off after correct refolding.”
As a different example, expression yields of pDNA, as high as 2 g/L, are based on BI’s E. coli host and a high cell density fermentation process. “A closed semi-continuous lysis system and purification process are used to manufacture large quantities of pDNA with high ccc content in a single downstream batch.”
For the future of biopharmaceutical contract manufacturing, Dr. Necina sees a demand for increased flexibility at the plant and reduced lead time, which means an increasing use of disposables and mobile equipment. “High-yield, high cell density fermentations will become more important in the future, challenging plant design and processes for primary recovery.”
“BI Austria is developing alternative downstream technologies such as crystallization and extraction. These technologies allow fast and cost-efficient processing of large amounts of biopharmaceuticals without investing in large vessels, expensive chromatographic resins, and chromatographic hardware. These processes can be operated as closed or continuous systems.”
High-yield Protein Expression Systems
ProBioGen (www.probiogen.com) and Evotec Technologies (www.evotec-technologies.com) recently completed their collaboration to develop methods and instrumentation for the selection of high-producer cells lines used in the production of biopharmaceuticals. The new system uses ProBioGen’s expertise in cGMP cell culture and Evotec’s Cytocon™ device.
ProBioGen has a high-yield protein expression system that is constructed through a combination of various optimized elements, such as promoters and selection markers. Added to the specific know-how of early clone separation and analysis, high-producer clones can be selected relatively early in the process to produce material for testing.
“Quality and capacity are prerequisites in this industry, but they do not guarantee business success,” Hing Kin Chan, Ph.D., vp commercial development at ProBioGen, says. “We also provide in-depth knowledge of structure-activity relationships when developing a cell line, the analytical methods, and the production process. This includes protein design, understanding the influence of different cell substrates on structure, pharmacology, glycosylation analysis, complex cell-based potency assays, and immunogenicity testing.”
ProBioGen is also developing its Membrane Based Bioreactor (MBB). “Our MBB provides the same benefits as a disposable system along with high cell densities,” explains Dr. Chan. “The MBB uses the same hardware for different volumes of bioreactors, and the first downstream step (clarification) is integrated in the upstream process, saving time and cost.” The company also has new membrane adsorbers in the downstream process for a higher flow rate and binding capacity.
“In the future, we will see an increasing use of disposable technologies, higher expression levels, and lower volumes of bioreactors used,” Dr. Chan notes. “We will have a flexible slots system for project allocation, parallel availability of perfusion and fed-batch in disposable bioreactors, and in-house MBB development. In addition, we are developing various new cell substrates for high protein expression, including an avian cell line that offers safe, reliable, easily scalable, and economical cell culture-based vaccine production. This cell line addresses the needs of the new vaccine types that are expected to dominate the market over the next 20 years.”
Streamlining Process Development
QSV Biologics (www.qsvbiologics.com) and Proteon Therapeutics (www.proteontherapeutics.com) signed an agreement for the manufacture of Phase I clinical trial material, a recombinant human protein produced from yeast. The yeast-based vasodilation drug will be used by patients with renal and vascular diseases.
QSV Biologics provides a variety of expression systems such as yeast-based and E.coli-based microbial fermentation, mammalian cell culture, and insect cell culture services. The company also recently signed an agreement with Tissue Therapies (www.tissuetherapies.com) for tech transfer and cGMP production of VitroGro®-1, a line of chimeric products using a single protein. “We will be using high-productivity cell culture manufacturing processes for these products,” Graeme Macaloney, QSV founder, says.
In addition, the company just signed an agreement to do a full process development and production project for clinical trial material for Seredigm.
QSV Biologics finds that its customers also require a strong ability in process development (PD). “To meet the evolving PD requirements, we are investigating bioprocessor systems that allow for rapid screening of bioreactor conditions and media optimizations,” explains Richard Hetrick, director of business development. “These miniature modeling systems could be extremely useful for streamlining the process-development bottleneck. In addition, we are evaluating some interesting chromatography products that eliminate gel-filtration polishing steps, which are inappropriate for industrial manufacturing.”
QSV Biologics has invested in disposables for holding and mixing vessels as well as disposable bioreactors for cell culture at the 50-L and 250-L sizes. In the next few months, QSV may serve as a beta site for a 1,000-L disposable bioreactor. “The benefits of using disposables for a CMO are obvious,” Hetrick says. Overall, we need to ensure that we meet our clients’ needs for being on time, on spec, and on budget without any regulatory issues.”
Goodwin Biotechnology (www.goodwinbio.com) uses either hollow-fiber or packed-bed perfusion bioreactors for its clients’ mammalian cell culture projects. “These reactors, though not thought by many to be scalable, are immensely productive, given their size and media feed,” Stephanie C. Finnegan, CEO, says. “We are adding large stirred tanks because that is what our customers want. However, these tanks are convertible tanks so they can also be run in perfusion mode. We believe that several events will occur in our industry: (1) titers will continue to improve; (2) targeted, or smaller-market therapeutics will become much more prevalent than blockbuster drugs; and (3) perfusion culture will become an accepted method for larger-volume production.
“Our clients come to us because of our experience in perfusion culture; none require huge quantities, and so they chose efficient modes of manufacture for smaller quantities. Now, with the larger bioreactors, we will continue to serve their needs as they progress to the market.”
Goodwin Biotechnology, in business for almost 15 years, serves a wide variety of international clients, including emerging and established biotechnology companies with Phase I to III products and government entities in early clinical development. “We act as consultants for our clients as well as economical and efficient expediters of products to and through clinical trials,” Finnegan says. “We are very aware of the Golden Rule of the CMO business: No two cell lines are alike, and each must be approached in development as a custom job. There will always be technologies that are appropriate for one cell line but not for another.”
Widespread Use of Disposables
Cytovance Biologics (www.cytovance.com) has a new custom-designed multiproduct facility for the production of therapeutic proteins and antibodies from mammalian cell culture. The company has been executing process-development projects for more than a year, and is now securing its first cGMP production contracts at the 100-L and 500-L scale, for both fed-batch and perfusion culture.
“We make widespread use of disposable technologies in buffer and media make-up, feeds, inoculum trains, harvests, and filtration,” Roger Lias, Ph.D., vp sales and business development, says. “In addition to the well-documented validation advantages, disposables allow us to employ flexible production scales for specific client needs, to rapidly turnover facilities between projects, and overall, to build a greenfield, state-of-the-art facility that meets the latest regulatory expectations.”
“Disposable bioreactor technology will become more significant as scale increases and mechanical systems supporting the technology become more robust,” Steven Perry, vp manufacturing operations, says. “On the downstream side, disposable membrane chromatography cartridges will increase processing speed and eliminate the column-packing operation. We continually monitor developments in single-use systems and will integrate new technologies into our offering as they become suitable for cGMP production.”
Cytovance is currently evaluating a number of production technologies on behalf of the developers on a contract basis, and will employ specific technologies if requested. “As a contract service provider, we do not want to be on the very cutting edge of new technology, unless specifically requested by a customer, because we do not want to introduce additional process risk that may compromise a customers’ continued development or regulatory submissions,” Dr. Lias asserts.
“Most of our customers are emerging and mid-sized companies seeking to avoid royalty burdens where possible. In addition, technology fit tends to be the more important differentiating factor when initially screening clinical-scale CMOs.”
Dr. Lias believes that quality and compliance requirements are non-negotiables in the biopharmaceutical contract manufacturing industry, and that capacity, availability, and flexible scheduling will continue to be important.
At the Baxter Bioscience (www.baxterbiopharmasolutions.com) Hayward facility, single-use containers from 1 L to 600 L are used for storage and transport of almost all buffers, media, process intermediates, and purified products. “Most recently, we have used HyClone’s (www.hyclone.com) single-use bioreactor (SUB) for process development and manufacturing of therapeutic proteins for Phase I/II clinical trials for several clients,” reports Kurt Kunas, Ph.D., senior research scientist at Baxter.
“As a single-use technology, the SUB has greatly reduced the amount of time, space, and expense that has traditionally been required for bioreactor capacity.” Because the HyClone SUB is a stirred-tank bioreactor, Baxter has been able to develop, scale-up, and transfer cell culture processes between SUBs and traditional bioreactors.
With bioreactors at the 250-L scale in the Baxter process-development laboratory, relatively large amounts of product can be generated for further downstream development without tying up the cGMP manufacturing suite. “In our process-development laboratory,” Dr. Kunas explains, “we currently operate two 50-L SUBs and two 250-L SUBs in addition to smaller-scale traditional screening bioreactors. We have successfully developed small-scale processes using traditional screening bioreactors, demonstrated scale-up of those processes using the SUBs in our laboratory, and then transferred the processes to the larger-scale traditional bioreactor suite. Our future plans include use of the 1,000-L SUB in the process-development laboratory and use of multiple 1,000-L SUBs in the cGMP-manufacturing SUB suite.”
Although capacity, compliance, and quality are the key success factors for contract manufacturers, and new technologies must be incorporated cautiously to avoid any risk to the product, most contractors are working steadily to increase the efficiency of their operation to save clients time and money.
Evolving biomanufacturing technologies, whether addressing the use of disposables, improved cell lines with high yields, high-throughput screening methods and modeling platforms for streamlined process development, optimized media, perfusion systems, and alternative downstream processing and purification methods, all contribute to the growing efficiencies in the biopharmaceutical contract manufacturing industry.
Sandra J. Fox is an analyst at HighTech Business Decisions, and William P. Downey is the managing director. HighTech Business Decisions published the industry study “Biopharmaceutical Contract Manufacturing: Improved Processes and New Capacity for Pipeline to Commercial Production,“ which is based on interviews with directors of biopharmaceutical manufacturing at 51 pharmaceutical and biotechnology companies, interviews with 33 contract manufacturers, and profiles of 97 contractors worldwide. Web: www.hightechdecisions.com. Phone: (925) 631-0920. E-mail: firstname.lastname@example.org.