Many technological advancements in recent years have enabled companies to shorten time to market, to better understand their manufacturing processes, and to characterize their products well. In BPI’s December 2013 issue (pages 47–50), I reported on the first half of an informal reader survey about those technologies, with commentary from some survey participants and others. This month concludes with my examination of analytical, formulation/fill–finish, and facilities technologies.Analytical Technologies
After writing several installments of our new “BPI Lab” series this year, I’ve learned a lot about the many and varied analytical methods used in biopharmaceutical quality, discovery, and development laboratories. But I needed to keep my multiple-choice lists under each survey question short because long lists can make people change their minds about participating halfway through! And there are so many technologies in this category, it was difficult to choose just a few. So I half expected the “other” category to carry the day here as knowledgeable readers filled in the blanks for me.
Surprisingly, the overwhelming winner wasn’t a technology per se — but rather a testing strategy that applies to many assay types. Nearly two-thirds of respondents said that “design of experiments (DoE)” was having the most impact on their analytical work. About a quarter highlighted improvements in liquid chromatography with mass spectrometry (LC–MS), and the remainder were divided between “-omics” approaches and a diverse “other” category. No matter what analytical method you’re using, it seems, DoE can help you optimize your results (24,25,26,27).
“Statistical DoE is a tool that is central to quality by design (QbD) and the development of product and process ‘design space’ (a combination of raw material and process variables that provide assurance that a quality product will be produced),” wrote consultant Ronald Snee for BPI in 2011 (25).
He went on to conclude that DoE’s elements, strengths, and limitations must be understood for it to be used effectively. “The promise of effective DoE is that the route of product and process development will speed up through more cost-effective experimentation, product improvement, and process optimization. Your ‘batting average’ will increase, and you will develop a competitive advantage in the process” (25).
Some survey participants were passionate about particular technologies. Among them, LC–MS carried the most weight (28). Proteomics and other “-omics” type studies (e.g., transcriptomics and secretome analysis) are also guiding characterization of products and cell lines, among other things (29). Honorable mention goes to automated cell counting, Factivity software, immunoassays, capillary electrophoresis (see the January 2014 installment of BPI Lab), analytical reverse-phase HPLC, and QbD in general.
“We are working with miniaturized immunoassays in a compact-disk format,” says Mats Inganäs at Gyros AB in Sweden. “The prime characteristics of this technology are automation, open system, short turn-around times, miniaturized reactions that reduce reagent and sample consumption, and efficient assay development.” So his company’s work seems to reflect much of what characterizes the most enabling analytical technologies in the 21st century.Formulation and Fill–Finish
Many biopharmaceutical companies focus entirely on producing an active pharmaceutical ingredient (API), then leaving drug-product manufacturing to specialized contract service providers (30). Quite a few major contract manufacturers do the same (31,32,33). For most people in upstream and downstream processing groups, formulation and fill–finish remain a “black-box” field. But preformulation best begins early in product development, and such characterization work can aid in process development. Similarly, fill–finish technologies such as lyophilization, spray-drying, and container–closures can make or break a product batch. In fact, many product failures come down to this crucial step in manufacturing — while the much larger number of successes go more or less unnoticed, of course, for business as usual.
And according to our survey results, that’s attributable to improved isolation technologies and automation of formulation and fill–finish operations. Those top two responses were close, but the latter edged ahead with 32% of responses over 29% for the former. Other contenders included prefilled syringes (12%) and other new container–closure systems (10%) as well as single-use technology. Companies need to take a balanced approach, as one small-company president located in North America put it, with costs, quality, and minimal waste as the important factors to weigh against one another. And whereas isolators help prevent contamination, automation works with them to reduce human error and add further process control. It would seem that the robots are fully ensconced into this aspect of the biopharmaceutical industry — even as they make in-roads with analytical laboratories and begin to show potential for upstream and downstream operations as well (34, 35). But like single-use technology and container–closures, they are not without their own challenges (36,37,38,39,40,41).
I asked Mario Philips (senior vice president and general manager for ATMI) for his thoughts on the subject. “The presence of pyrogens in parenteral products is a real patient risk,” he pointed out. “If these substances are introduced into the bloodstream, they can rapidly induce fever and even cause sepsis or progressive septic shock. Using guaranteed sterile and pyrogen-free vials is one way to prevent that from happening. Vials would otherwise have to be depyrogenated on site before use, which generally involves heat treatment to destroy residual pyrogens. Pyrofree vials, for example, are thoroughly depyrogenated before shipping to ensure at least a three-log reduction in pyrogens. They are then double-vacuum-packed in a polyether ether ketone bag to maintain their pyrogen-free status.
After making great inroads with biopharmaceutical processing, he said, disposables are now being applied to fill–finish. “One main advantage is the huge reduction in validation requirements. Equipment is used only once, saving time and resources otherwise spent on cleaning and associated testing. The disposable platform being commercialized by ATMI and Disposable-Lab is similar to traditional fill–finish equipment but on a much smaller scale. Connectors, vials, dispensing needles, baskets, and caps are all disposable, and unavoidably nondisposable components are decontaminated with hydrogen peroxide before use. Filling takes place in an isolator.”
What About Discovery?
We at BioProcess International won’t pretend to be experts in drug discovery — or even amateurs in the classical sense of the word. Our interest lies in drug development, all the work that follows selection of a worthy candidate. However, the curiosity factor drove us to ask survey participants about how technological advancements have improved the speed and efficiency of discovery efforts. In particular, we wondered whether the human genome project was yet beginning to pay off — and whether all the talk of systems biology and buzzwords
that end in “-omics” could be more than just hype.
The most powerful approach would seem to be “-omics,” topping out our survey results with a combined score of 42% of respondents checking either the “-omics” (31%) or “human genome project” (11%) box. Close behind is systems biology with 38% of results. Finally, 22% of respondents mentioned “other” specific technologies such as high-throughput screening and antibody fragments.
There is definitely some overlap here. Obviously the human genome project is part of the “-omics” revolution. But genomics, lipidomics, metabolomics, proteomics, and transcriptomics are all contributing to systems biology as well. And none of them would be possible without high-throughput screening/sequencing techniques. Modern antibody fragment proteins are but one result of all this work.
In a recent blog entry, Alexander Kamb and Sean Harper (both vice presidents at Amgen) discussed the impact of the human genome project on drug discovery (1). They focused on
genome-wide association studies (“nearly a bust in terms of delivering actionable information for drug discovery” after providing many hints about disease-relevant genes)
ultra–high-throughput sequencing (for “comparisons at maximum sensitivity across thousands and, in due time, millions of genomes from individual people”)
searching for single-nucleotyped polymorphisms and rare variants that “can be much more informative for guiding development of new medicines.”
Ultimately, Kamb and Harper called the human genome “nature’s gift to medical science. Without it, drug discovery would remain at the mercy of disease models in animals, and drug development would remain a high-risk enterprise marked by frequent, expensive failures caused by our limited knowledge of valid human disease targets.”
In 2001, Thomas Reiss of the Fraunhofer Institute for Systems and Innovation Research in Germany predicted that the published human genome could increase the number of drug targets by “at least one order of magnitude” (2). He also cautioned that it would require new approaches to drug discovery, particularly involving cooperation between biotechnology companies, big pharma, and research instituations. We are seeing that change now — even if the results have yet to provide a flood of new classical and biologic drugs. And from what we know of drug-development timelines, even a target coming immediately out of the first genome publication would only now be finding its way into the clinic. So I believe that the best is yet to come.
1 Kamb A, Harper S. The Human Genome Project Wasn’t Overhyped; The Payoff Just Took Time. Xconomy.com 3 October 2013.
2 Reiss T. Drug Discovery of the Future: The Implications of the Human Genome Project. Trends Biotechnol. 19(12) 2001: 496–499.Facilities
Biomanufacturing plants are undergoing a 21st-century revolution as they transform from massive, hard-piped factories devoted to making single, high-volume products into flexible, multiproduct facilities designed with an uncertain future in mind. Many of the big biotech products that are beginning to lose their patent protection have been made using 20th-century processes that rely on stainless steel equipment and utility piping — with complex steam-in-place/clean-in-place (SIP/CIP) systems making up a substantial portion of the latter. Cleanrooms, controlled air-flow, and associated staff training and gowning have been at the heart of contamination control. Biosimilar competitors in development will be made in 21st-century facilities based on single-use and isolation technologies — without cleaning validation associated with SIP/CIP processes but with the added burden of leachables and extractables validation instead.
How new plants are built has also changed. Modular approaches speed up the process (42, 43), but so-called “ballroom” designs (wherein upstream and downstream areas are simply big rooms in which closed, single-use systems can be connected, disconnected, and wheeled about like dancers on a dance floor) may make them unnecessary. That may be the case, as our survey results suggest: Only 9% of participants said that modular construction was the most enabling technology for biofacilities. Another 9% chose “other” and suggested more specific developments such as in-line buffer dilution and sandwich paneling from French company Dagard.
I was pleased to see 21% of respondents choosing to highlight sustainability solutions such as solar power and other alternative energy sources, water and energy conservation, and process intensification (44). Environmental considerations are another aspect of 21st-century facility design — brought into light by the energy crisis and associated cost concerns — and could provide one means for the drug industry to counter the “bad PR” associated with pricing and product development timelines (44, 45). Notably, more European survey participants than North American respondents pointed to sustainability as an enabling technology. And in many cases, it is suppliers of products and services to the bioprocess industry that are leading the way (e.g., DSM Biologics, EMD Millipore, NewAge Industries, Pall Corporation, and Vetter Pharma International).
Environmental concerns were initially brought up when single-use technology began to see wider adoption for bioprocessing (43, 46,47,48,49,50). But most companies have found that significant reductions in energy and water use along with elimination of many caustic cleaners have outweighed the increase in consumables associated with disposable components. In fact, disposal options themselves have diversified toinclude incineration for energy production among other possibilities.
This brings us to the clear winner in facilities: 61% of our survey participants said that single-use technologies were having the greatest impact on their companies’ approach to facilities. And this makes sense. Whether combined with stainless steel in hybrid, modular plants or filling ballrooms with plastic, disposables are changing the face of bioprocessing (51,52,53).
I asked Jean-Louis Weissenbach (R&D director of single-use processing systems EMD Millipore) about these results. “The key challenges in biopharmaceutical manufacturing,” he pointed out, “have always been cost of goods (CoG), speed to market, and increasing regulation. Most production facilities were and are still using conventional stainless steel equipment, including hard-piped installations throughout upstream and downstream operations. Such equipment is relatively inflexible and requires significant capital investment. Further, the layout of the facility itself is dictated by these conventional installations.”
Weissenbach says that cost factors have pushed the industry to think differently. “Single-use technology continues to gain a solid foothold in the market. Cleaning between manufacturing runs is key in a CGMP environment. The shift toward single-use technologies presents significant advantages — e.g., reducing cleaning requirements and cross-contamination risk — while increasing flexibility and speeding turnaround between product campaigns.” Set-up and facility use are improved as well, as the need for steam generation is reduced.
“This past decade has seen increased development and adoption of single-use technologies,” Weissenbach concluded, “an
d the trend is expected to continue as scale and standardization challenges are addressed.”
Eva Lindskog, PhD (upstream marketing segment leader at GE Healthcare Life Sciences) also sees this as a true paradigm shift. She told me that the industry is currently in transition, with single-use technology maturing to a point where it supports more demanding applications and starting to become a truly integrated part of the overall business strategy for biopharmaceutical companies. “For example, recent developments in single-use bioreactor technology now enable the use of disposables throughout the whole drug development process,” she said, “from preclinical process development to 2,000-L manufacturing scale. In our experience, single-use technologies are being used in a range of processes beyond conventional CHO MAb production: for example, in microbial fermentations, vaccine manufacture, and immunotherapeutic applications. We see pipeless plants being constructed with a design that is entirely based on single-use equipment.”
Lindskog points to several factors driving wider use of disposables: higher product titers, fewer blockbuster products, increased competition, and increasing cost awareness. “Single-use technologies offer the potential for reduced overall production costs,” she explained, “by decreasing upfront investment and shifting a bigger portion of the overall cost to just-in-time.”Best in Show
Taken as a whole, our survey results point to one unavoidable conclusion. It probably comes as no surprise that single-use technologies top the long list of advancements and inventions that are enabling the maturation of the biopharmaceutical industry. They have drastically changed seed trains and production suites and are having incremental impacts downstream while promising radical transformations of filling lines and facilities overall. Disposables are eliminating some analytical work (e.g., cleaning validation) while replacing it with other needed testing (e.g., extractables and leachables). They are making multiproduct facilities more workable. And single-use technology has put a new image of bioprocessing in our heads: Once dominated by shining stainless steel, it’s now just as likely to involve glowing polymers.
Most of the trends identified herein are continuing. Meanwhile new technologies seek to disrupt the status quo. It’s fair to say that the biopharmaceutical industry isn’t just maturing; it’s matured. And as much as it is based on science and biotechnology, these advancements have made that possible. New ones will join them in keeping the ball rolling. Innovation is as much a key to success in bioprocessing as is regulatory compliance. Companies that fail to keep up with the advancing technology will be left behind in the race to market.
Cheryl Scott is cofounder and senior technical editor of BioProcess International, 1574 Coburg Road #242, Eugene, OR 97401; 1-646-957-8879;