For small companies seeking to produce biopharmaceuticals or vaccines, the capital expenditures required are often daunting. Financing is especially difficult to obtain in the current economy. To reduce costs, some small companies and contract manufacturers have turned away from fixed equipment and adopted flexible manufacturing systems. These companies have discovered that flexible manufacturing also has benefits for production and regulatory compliance. Large biopharmaceutical manufacturers are beginning to take notice. Could flexible manufacturing change the standards for biopharmaceutical production? To find out, 

 talked to James Robinson, vice-president of technical and quality operations at biotechnology company Novavax (Rockville, MD).  

EPR: How do you define “flexible manufacturing?”  

Robinson: When we use the term “flexible manufacturing,” we refer to single-use systems that can be rearranged for multiple manufacturing programs. Single-use systems are generally portable when not full of product, and such unit operations can be reorganized based on a specific product need. Reorganization is challenging with fixed equipment and stainless-steel piping networks.  

We also use the term “modular manufacturing” at Novavax, which refers to the concept of designing, qualifying, and validating a production module—including all unit operations necessary to produce a product—once and duplicating it in multiple facilities or locations.  

EPR: What are the production advantages of a flexible manufacturing facility for the biopharmaceutical industry?  

Robinson: With fixed equipment, each facility and its supporting infrastructure are unique, and it is difficult, if not impossible, to make exact copies. Flexible equipment allows us to expand capacity by adding cell-culture modules for increased throughput without having to reengineer the entire process. We can design a facility that manufactures 50 million doses of influenza per year with one or two reactors supported by a single downstream process train. Adding two reactors allows the capacity to be doubled with only the added cost of the reactors. Capacity is further increased by duplicating the entire production module.  

If a process can be designed to use only single-use systems, a significant amount of facility infrastructure can be avoided. Fixed equipment requires miles of piping and hundreds of valves and process controls to maintain the cleanliness and sterility of equipment for reuse. Flexible manufacturing systems allow a company to shrink the plant and its cost by using less equipment, piping, valves, and controls. They focus the company’s investment on the manufacturing equipment itself.  

Reduction in facility complexity speeds the design and validation of a facility. Reduction in complexity also greatly reduces the risk of project delay and cost overruns. When Novavax commissioned a conceptual facility design for an influenza-manufacturing plant, we were given an estimated cost of $40 million to support a 75-million-dose/year bulk-manufacturing facility. The two newest facilities in the US, which perform at a similar scale, are reported to have cost $150 million and $600 million. In addition to the lower cost, we predict a project timeline of less than two years, compared with the four- to five-year plans for these other facilities.  

EPR: When would a flexible manufacturing facility be undesirable?  

Robinson: Some processes are not supported by single-use systems today. If you require a homogenization step or strong solvents, for example, or if the process is low-yield, and therefore high-volume, the systems could be limiting. In addition, if your process cannot be 100% single-use, your company will need to invest in the added facility infrastructure—such as clean-in-place and sterilize-in-place systems—and the longer validation times associated with traditional plants. You can minimize the impact of these systems and still gain the lower level of complexity in many of your systems, but not the full benefit of the financial or time savings.  

EPR: Can you offer a specific example of a flexible manufacturing operation in effect from your own work in the industry?  

Robinson: Novavax has developed a 100% single-use process for the manufacture of its influenza vaccine using recombinant viruslike particles. The result is that we can build a facility for a fraction of the cost and in half the time of a traditional flu facility. We can do this because of the high yield and low biosafety level of our baculovirus manufacturing platform. We have been able to avoid processes that are not available in a single-use format such as ultracentrifugation, which is often used in egg-based influenza-vaccine production.  

EPR: Is flexible manufacturing limited in terms of scale?  

Robinson: The largest bioreactor available today holds 2000 L. In about five years, we expect 5000-L reactors to be available. This scale would satisfy the majority of vaccine-manufacturing processes that exist today. The downstream technology is a little more limited as volumes increase, although new technologies now are being developed for moving and processing high flow rates with single-use systems.  

EPR: Do you foresee Big Pharma adopting flexible manufacturing in the near future?  

Robinson: Big Pharma has been slow to adopt these technologies so far. These companies’ traditional infrastructure is already in place and validated, and many manufacturers believe that the cost of changing processes would be higher than continuing to use their current equipment. The new technology also may seem risky to some big companies.  

I believe that the value gained in time savings for new capacity and the flexibility of this technology for multipurpose facilities will incite organizations to adopt these technologies over the next five to 10 years as new products replace existing products in their portfolios. 

Articles

Production Lines

Could flexible manufacturing change the standards for biopharmaceutical production? To find out, Equipment and Processing Report talked to James Robinson, vice-president of technical and quality operations at biotechnology company Novavax (Rockville, MD).

Biopharmaceutical Manufacturers Look for Flexibility

 translation systems are based on the principle that protein synthesis does not require an intact cell. This principle led to the development of cell-free protein synthesis by Zubay in 1967. In cell-free production systems, translation is accomplished using a crude cell lysate that provides the necessary translational machinery, accessory enzymes, transfer RNA, and translation factors. The process also requires the addition of a nucleic-acid template, amino acids, and an energy source.

The primary limitation of cell-free, batch-mode reactions is their poor productivity, which results from the transience of the primary energy sources. Phosphoenolpyruvate (PEP), a phosphate energy source, is depleted rapidly, even in the absence of protein synthesis (1). In 1988, Alexander Spirin and colleagues at the Institute of Protein Research in Moscow created a continuous-flow, cell-free translation system to address the problem. This system continuously removes reaction byproducts and relies on a continuous supply of energy and substrates (2). Despite its improvement in yield, however, this system’s operational complexities make it extremely impractical.

By replacing PEP with 3-phosphoglycerate as the primary energy source, productivity can be doubled. The latter energy source enables a steady supply of adenosine triphosphate (ATP), which stimulates protein synthesis (3). ATP production is sustained for a longer time in the presence of 3-phosphoglycerate than it is in the presence of PEP (4). Optimizing the reaction conditions in buffers by adding stabilizers for enzymes and improving the incubation conditions can improve productivity even further (5).

Several laboratories have tried to develop high-throughput, cell-free protein-synthesis systems (6–8). Swartz demonstrated that by using efficient and economical alternatives to PEP (e.g., pyruvate), mimicking 

’s cytoplasmic environment, and stabilizing amino acids, it is possible to produce nearly one milligram of protein per milliliter of reaction using a batch format (9, 10).

The current authors sought to improve the productivity of protein synthesis by using a highly active cell-free extract from 

 and by optimizing buffer conditions and shaking conditions. We also designed a new and simple device for high-throughput reactions. Using common laboratory equipment and materials, our system produces as much as 4 mg of green fluorescent protein (GFP) per milliliter of reaction in high-throughput and large-scale formats. This yield is equivalent to 4 g of GFP protein per liter of reaction (see Figure 1).

A high-throughput reaction requires a 48-well tissue-culture plate and 48 dialysis units. The authors performed cell-free reactions in quantities as large as 5 mL using a covered plastic container, a laboratory dialysis membrane, and clamps. Figure 1 demonstrates the similarity between the productivity of high-throughput (50-µL) and the large-scale (5-mL) operations. The system was about 10 times more efficient than conventional batch reactions with respect to the yield of GFP.

The technology could be useful to manufacture pharmaceutically important proteins on a much larger scale. A large-scale reaction device could be made by applying the techniques for scaling up standard dialysis reactions. Large-scale production of 

 extract might result in a bottleneck, but it could be overcome by large-scale or continuous fermentation, centrifugation, and cell extraction.

	K. Sitaraman and D.K. Chatterjee, “Energetics in 

 Kudlicki et al., Eds. (Landes Bioscience, Austin, TX, 1st ed., 2007), pp. 42–52. 

	T. Kigawa et al., “Bacterial Cell-Free System for Highly Efficient Protein Synthesis,” in 

 A.S. Spirin and J.R. Swartz, Eds. (Wiley-VCH Verlag, Weinheim, Germany, 1st ed., 2008), pp. 83–97.  

Takashi Yabuki is a visiting scientist, and Deb K. Chatterjee is an associate director and senior principal scientist, both at SAIC-Frederick, the National Cancer Institute at Frederick, Frederick, MD 21702, 

In the Mixer

The authors sought to improve the productivity of protein synthesis by using a highly active cell-free extract from Escherichia coli and by optimizing buffer conditions and shaking conditions.

Large-Scale Protein Production with a Cell-Free System

Malvern
 Software facilitates chromatographic measurements


 Malvern’s (Malvern, England) OmniSEC software supports gel-permeation chromatography (GPC) and size-exclusion chromatography (SEC) in quality-assurance applications. It enables proteins’ absolute molecular weight to be measured accurately with light scattering. The product also can determine the solution’s concentration and oligomeric composition. In addition, the software displays the size of proteins, thus enabling scientists to check for denaturing, oligomers, aggregates, and contaminants.

The product’s polymer-branching view helps indicate the density of drug-delivery matrices. Scientists can use the view to determine whether an active ingredient loads equally in the matrix at different weights.

The system simplifies GPC and SEC by selecting the baseline and integration limits automatically. Scientists can obtain results with two mouse clicks and do not need to adjust the software during operation. The product can be used for lot release at production sites.

OYSTAR USA Process Division
 Machine performs efficient granulations and particle coating


The Hüttlin HDGC fluid bed from the OYSTAR USA Process Division (Fairfield, NJ) allows operators to perform fluid-bed drying, granulation, and coating in one product container. The device’s Diskjet air-distribution plate includes laser-cut, 150-µm wide slots positioned radially at a 45° angle. The slots enable process air to move product upward in a circular motion, unlike the convection movement in a typical fluid-bed dryer. The slots also move the product at a higher velocity than a typical unit does. The circular motion and higher speed facilitate fast and homogeneous mixing.

The Hüttlin HDGC machine uses the same nozzles, which are embedded in the Diskjet at a 45° angle, for bottom-spray coating and granulation. The nozzles include a microclimate compressed-air supply that helps prevent nozzle blockage. Because of the circular motion, product passes in front of the nozzles more frequently than it does in a typical fluid-bed dryer, thus reducing granulation time by about 30%, according to the company.

PharmTech's monthly newsletter, Equipment & Processing Report, reviews the Editor's Picks for the January 2010 edition from Malvern and OYSTAR.

Equipment and Processing Report-01-20-2010

Biopharmaceutical Manufacturers Look for Flexibility

Large-Scale Protein Production with a Cell-Free System

January 2010 Editor's Picks: Products from Malvern and OYSTAR