Two recent industry developments—Genzyme's delay in gaining approval for its commercial-scale manufacturing of a new drug, and Amgen's announced layoffs—raise concerns about biotech performance at large scale. In the first case, the issue is manufacturing scale-up; in the second, it's how companies sustain long-term success once they become big. But in both cases, industry pundits are extrapolating conclusions from these issues that the facts don't seem to support.  

, Genzyme has been unable to get FDA approval for its large-scale production of Myozyme, a drug approved last year for Pompe's disease. According to the article, FDA says the product manufactured in the large-scale reactors at the company's Boston facility is not the same as the product used in clinical trials, produced in smaller equipment at the company's site in Framingham, MA. The product manufactured in Boston is approved for sale in 29 other countries, however, and Genzyme maintains it is safe and effective.  

Biologics manufacturers are quite familiar with the challenges of scale-up, so this case, though disappointing, is not unique. New this time, however, is how this case is being tied to the debate about follow-on biologics. In a 

blog, journalist David Amstrong says the fact that an experienced company like Genzyme is having trouble manufacturing its own product heightens concerns about the ability of generic manufacturers to copy biotech drugs. 

But regulatory measures can handle situations like this. Companies regularly file comparability protocols to demonstrate product equivalence following manufacturing improvements or after transferring operations to another site, and Genzyme can use such testing in this case. In the same way, if a biosimilars law is passed in the US, nonclinical testing could be used to support the equivalence of follow-on biologics. In all cases, if such testing appears insufficient, clinical trials should be required. So scale-up challenges like Genzyme's should not be used to raise the spectre of unsafe biosimilars reaching the market. 

The other current industry development is Amgen's announcement that it will lay off up to 2,600 workers, following a significant decline in Aranesp sales resulting from safety concerns and reimbursement changes this year. Some financial analysts have said the cause is that Amgen has become too big. It is no longer nimble, they say, and thus is facing the same kinds of problems that Big Pharma has faced—reliance on blockbuster products; decreased R&D productivity; difficulties restocking its pipeline.  

It's easy to conclude that Amgen's current challenges are because of the company's size. But, sadly, drug safety issues can happen to any company, large or small. If the same happens to an emerging biotech firm, no one on Wall Street says, "well, they just got too big."  

So in analyzing the industry's current challenges, let's be careful before extrapolating about what they mean.

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In analyzing the industry's current challenges, let's be careful before extrapolating about what they mean.

From the Editor: Issues of Scale

If there really can be "too much of a good thing," providers of contract development services may be experiencing it. Contractors are having to admit that the robust demand for services arising from the surging new drug pipeline may be pushing them to the limits of their ability to deliver.  

That's a key insight gleaned from the 2007 survey of market conditions conducted by PharmSource in conjunction with 

. The survey was conducted in May and received 544 responses, including 432 from biopharmaceutical companies and 112 from service providers. All respondents were involved in buying or providing chemistry, manufacturing, and controls (CMC) services, including analytical chemistry and microbiology, formulation, process development, packaging, and API and dose manufacturing 

Service providers themselves report that rapid growth is causing strains. Nearly 55% of CROs admit that lack of capacity is constraining their ability to take on new business; that's up from 43% in 2006. Customers have noticed how busy their suppliers are: in this year's survey, 30% say that contractors appear to have all the work they can handle; up from just 13% in 2006. 

Interestingly, staffing is not the biggest problem facing service providers; rather, it's their ability to manage the growth. The biggest single constraint cited by service providers is organizational processes, with inadequate capital to fund growth a close second. When asked whether their management and systems were adequate to handle growth of 20% or more, only one-third indicated that their systems and management are up to the task (Figure 1). A quarter of contractor respondents admit that neither their systems nor management are adequate to handle current growth, while the rest concede that one or the other is lacking.  

The capacity problems reflect an unprecedented level of activity in the development services industry. More than a quarter of service providers expect revenues to grow by 20% or more this year; only 15% of contractor respondents expected such robust results in last year's survey. The growth is coming from all customer segments, with mid-size and generic bio/pharmaceutical companies showing particularly big jumps.  

The contractors' success is mirrored in the responses from bio/pharmaceutical company professionals, 94% of whom report an increase in spending on outsourced activity for the year. The survey results indicate that the growth reflects a fundamental move in favor of outsourcing rather than doing things in-house. Among bio/pharmaceutical company respondents, 27% reported that their spending on contract services is growing faster than their overall spending, up from just 18% of respondents in last year's survey. Over 20% of respondents report that half or more of the spending in their particular area is outsourced; that's more than double the response from 2006.  

The increase in spending is remarkably consistent across company size, and rates of growth for spending on commercial and development services are also about the same. 

The growth in spending has created a more fluid contract services market that offers more opportunities for service providers. Among bio/pharmaceutical company respondents, 57% report that they are actively looking for new service providers, either to increase their vendor base or to replace current providers; that's up from 37% in 2006. Only 5% of respondents say they are sticking just with current vendors. 

That search for new vendors may also reflect growing dissatisfaction with current vendor relationships. The survey suggests that service providers do not fully appreciate how their growing pains are impacting their customers.  

When asked how they think their clients would rate the quality of their customer service, 36% of suppliers say customers would rate them as "excellent" (Figure 2). However, when customers are asked how they rate their service providers' customer service, only 6% rate them as "excellent," and vendor performance on project management and technical/operational dimensions got similar marks. Although  customer ratings of their service providers are not dire—most rate their vendors as "good" or "satisfactory" and less than 20% rate them as "fair or "poor"—the  survey results suggest that there is a big gap between customer and service provider definitions of service excellence.  

In fact, meeting service levels is a major challenge for pharmaceutical services providers because the requirements of their client base vary widely. While major bio/pharmaceutical companies will judge service providers on the basis of technical and operational performance, small bio/pharmaceutical companies expect help in navigating the drug development process and may not appreciate technical and operational excellence. Further, service providers have yet to find an effective way of demonstrating superior customer service and project management capabilities, and if they can't put a value on it, they can't justify a price premium for it. 

Service quality issues might intensify next year if our respondents' expectations for 2008 come to pass. Among bio/pharmaceutical company respondents, 43% expect spending for contract services to grow by 10% or more next year (versus 36% in last year's survey), and only 7% expect spending to decrease. Service providers are very optimistic as well: 80% expect 2008 to be better or much better than 2007. In 2006, only 55% had such expectations for the following year. 

Further growth threatens to intensify the pharmaceutical services industry's capacity and service challenges. The industry continues to be dominated by small companies whose technically inclined owner–operators may lack the inclination and executive skills to deal with the business issues of their rapidly growing enterprises. An accelerating process of consolidation promises increased scale and greater management sophistication, but for now even the largest players seem to be struggling to meet the mounting expectations of customers. Still, it's unlikely that anyone is going to say "no" to new business while they work out their capacity issues.  

Of course, the market situation could change quickly. Venture capital—a major source of the funds that ultimately flow to CROs—can dry up quite suddenly and whole therapeutic classes can be discredited. With major pharmaceutical companies undergoing rounds of cost-cutting and unfavorable reimbursement and patent trends, a darkening outlook could alter the landscape for service providers in very short order. 

 is president of PharmSource Information Services, Inc., Springfield, VA , 703.383.4903, 

Meeting service levels is a major challenge for pharmaceutical services providers because the requirements of their client base vary widely.

Outsourcing: Robust Market Testing CROs' Ability to Deliver

Legislation to reauthorize the Prescription Drug User Fee Act (PDUFA IV) will provide more tools and legal authority for the US Food and Drug Administration to monitor and mitigate drug risks, while also boosting user fees and extending several FDA programs. The bill, which the Congress is expected to finalize by the end of this month, implements an FDA–industry user fee agreement issued in January, and a similar plan for medical devices. Additional provisions renew incentives for manufacturers to study pediatric uses of drugs and establishes such incentives for medical devices.  

To strengthen drug safety policies, the bill gives FDA authority to require postapproval clinical trials and to revise product labels within a set time-frame. Manufacturers will have to disclose more information about ongoing clinical trials and study results. There is more funding for FDA oversight of postmarket drug use and to expand the agency's information systems for tracking adverse events and for detecting emerging safety problems. Also, manufacturers of medications known to be risky will have to spell out a range of pharmacovigilance activities to inform patients and prescribers of possible dangers and to ensure appropriate product use. These initiatives should provide more timely information on emerging concerns about certain medicines, but also could drive up the cost of developing new therapies and delay access to new treatments.  

In adopting PDUFA IV, Congress will boost the annual fees paid by pharmaceutical and biotech companies to adjust for inflation and the increased cost of maintaining an efficient and timely drug development and review process. FDA and industry initially agreed on a new user fee program that would collect nearly $400 million a year starting in 2008, with steady increases over the program's five-year period. An additional fee program bolsters FDA review of drug TV advertising further. 

The legislators increased drug user fees even more to support a number of new safety initiatives. An important change is to authorize the use of fee revenues for drug safety oversight and assessment throughout a product's lifecycle—and not just during the first two or three years after product approval, as currently allowed. Moreover, the bill gives FDA authority to require a risk evaluation and mitigation strategy (REMS) from companies when the regulators decide that a formal approach is needed to manage particularly risky products. A REMS would evaluate the need to: 					

conduct additional postapproval studies and clinical trials to assess a specific safety signal

require labeling changes to disclose new safety concerns

prepare and distribute a medication guide or patient package insert

develop a communication plan for disseminating additional healthcare information to healthcare professionals

seek FDA pre-review of advertising and marketing materials to ensure that ads describe serious risks appropriately and fairly

limit drug prescribing and distribution. FDA may determine that specific treatments should be prescribed only by specially trained health professionals and for certain patients. Distribution similarly may be limited to select wholesalers and pharmacists. Such limited access programs may require additional laboratory testing, patient monitoring, and patient enrollment in a registry. 

In addition to establishing a REMS for certain products, the legislation enhances FDA authority to implement new oversight and disclosure policies for all drugs. These include: 					

: Congress strengthens FDA authority to require manufacturers to make certain labeling changes to reflect new safety information. Instead of  lengthy discussions between sponsors and FDA about the need to revise a product label, the legislation sets timeframes for negotiating such changes. The new system offers a process for sponsors to challenge a requested labeling change and for resolving disputes. 

Timely completion of postapproval studies

: FDA currently may request additional clinical trials and analyses following approval of a new therapy, but lacks authority to penalize companies that fail to complete these studies. The new policy calls for discussion of postapproval study options earlier in the approval process to ensure that agreements are realistic and address important issues. Manufacturers will have to propose timetables for completing these studies, report periodically on their progress, and explain why they cannot meet the set goals if problems arise.

More information on active clinical trials

: To better inform patients of opportunities to participate in clinical trials, sponsors will have to submit information about more ongoing studies to the 

 website. The legislation expands listings to cover most regulated drugs and biotech therapies. It also beefs up registration listings to include a long list of specific information, including whether there is any expanded access program for a not-yet-approved test treatment. 

: A key goal of the new policy is to link clinical trials registration to results information. Many manufacturers already post study results voluntarily, but the legislators seek a more comprehensive approach. Policymakers expect that such listings will include published studies and all FDA information on the drug (except proprietary trade secrets), such as the approved product label and relevant safety information. 

: The legislation reflects Congressional concerns that too many members of FDA advisory committees have financial links to the industry. Although policymakers recognize that these panels can benefit from input from experts who may have such conflicts, the legislation aims to make advisory committees more objective on safety issues by limiting waivers needed to permit such testimony. 

A prime initiative for reducing drug safety problems is to modernize FDA's electronic information systems. User fees have supported improvements in the agency's IT infrastructure for new drug review and oversight; now PDUFA IV will provide added funds to establish an all-electronic regulatory submission and review environment over the next five years. The envisioned system will allow manufacturers to send in applications electronically with automated cross-links to previously submitted data, while FDA reviewers will be able to retrieve all relevant submissions and data and  tools for searching and analyzing them. The goal is a system that can handle two-way transmission of regulatory correspondence.  

A modern IT system also would improve FDA's current adverse event reporting system (AERS) for collecting and analyzing such reports from manufacturers, health professionals, and patients. In addition, Congress supports FDA efforts to build a more robust active surveillance system instead of relying on spontaneous adverse event reports to identify emerging safety problems. Additional resources will permit FDA to contract with government health programs and private health plans to access population-based epidemiological data on millions of patients to expand postmarketing drug surveillance. More collaboration with government and academic organizations and with the Centers for Education and Research on Therapeutics (CERTS) will enable FDA to learn more about how a treatment works in real-world postmarket circumstances, including the extent and outcomes of off-label drug uses.  

PDUFA funds also will expand FDA staffing of epidemiologists, safety evaluators, and programmers who are able to use such IT resources. New research in this area should provide a better understanding of the role of epidemiologic analysis and use of databases in evaluating medical product safety. 

's Washington editor, Chevy Chase, MD, 301.656.4634, 

The new legislation authorizes the use of fee revenues for drug safety oversight and assessment throughout a product's lifecycle.

FDA Leglislation Tackles Safety Issues

 The year 2006 witnessed the approval of 13 biopharmaceuticals in the United States or European Union (Table 1) including four hormones, three antibody-based products, three therapeutic enzymes, one recombinant vaccine, an anticlotting agent, and a nucleic acid–based product. Major target indications included hereditary genetic conditions (four products, three of which—Elaprase, Naglazyme, and Myozyme—have orphan status), growth deficiency (two products), and neovascular macular degeneration (two products).  

 Table 1. Biopharmaceuticals (recombinant proteins and monoclonal antibody-based products) approved in the  US or EU in 2006   			

Only 10 of the 13 products were genuinely new to the market. Naglazyme, Tysabri, and Macugen, although approved in one region last year, had gained approval before 2006 in some other world region.    

In terms of expression systems used, five of the approved biopharmaceuticals are produced in mammalian cell lines, four in 

 again confirming the prominence of these three expression systems in the biopharmaceutical sector. One product (Macugen) is produced by solid-phase organic synthesis. Another (Atryn) is particularly noteworthy because it is the first biopharmaceutical produced in a transgenic animal to gain regulatory approval. 

Like 2005, last year also witnessed notable approvals in the context of nonparenteral delivery systems and biosimiliars. The approval of Exubera (a recombinant insulin) marks a watershed for pulmonary-based biopharmaceutical delivery. The biosimiliar product Omnitrope (recombinant human growth hormone), approved in 2005 by the Australian authorities, gained approval in both Europe and the US in 2006. And the EU further underlined its acceptance of the principle of biosimilarity by approving a second product (Valtropin, also a recombinant human growth hormone) last year.  

The remainder of this article focuses on individual products approved in 2006, with product information detailed in a monograph format. Information was drawn from regulatory sources

 and the Internet homepages of  sponsoring companies. Because monographs detailing  Naglazyme and Macugen were included in previous articles

 they are not considered below. Although Tysabri also has been covered before

 , a revised monograph is included here because of the altered indications and restrictions linked to its resumed approval last year.  

 (antithrombin alfa) is a recombinant form of human antithrombin (AT) produced by transgenic goats in their milk. The 432 amino acid single-chain protein is, like the native human molecule, glycosylated. It harbors four N-linked glycosylation sites (Asn 96, 135, 155, and 192) and three disulfide linkages. The recombinant molecule displays significant differences in its glycosylation profile compared with native human AT, both in terms of monosaccharide composition (for example, it is fucosylated and less sialylated) and overall side chain structure. This influences the pharmacokinetics and heparin binding affinity of the product. 

The European Commission approved Atryn under exceptional circumstances in July 2006 because the rarity of the condition prevented generation of comprehensive clinical data. The product is an anticlotting agent and is indicated for the prophylaxis of venous thromboembolism in surgery of patients with congenital antithrombin deficiency. The original marketing authorization holder was Genzyme Europe, but this has since been transferred to Leo Pharma (Ballerup, Denmark).  

Native AT is a plasma glycoprotein that is synthesized and stored in the vascular endothelium and normally present in blood at levels of 125 Âµg/mL. It is one of the most significant natural inhibitors of blood coagulation and brings about its anticlotting effect by complexing with, and hence directly inhibiting, several blood clotting factors, most notably thrombin (factor IIa) and factor Xa. The complex is then rapidly cleared from circulation via internalization and degradation in liver cells. In addition to a coagulation factor binding domain, AT displays a heparin binding domain; binding of heparin greatly accelerates the rate of complex formation. Heparin is normally co-administered with the product. 

Hereditary AT deficiency can be categorized by a decrease in absolute serum AT levels or by the presence of mutated (dysfunctional) forms. The major clinical consequence is an increase in the risk of clot formation in deep veins, pulmonary embolism, and venous clot formation at uncommon sites (e.g., hepatic, retinal, or cerebral venous thromboembolic events). Particularly high-risk circumstances include surgery, bed rest, and pregnancy. 

The manufacture of Atryn involves initial harvest (i.e., milking the animals), followed by clarification and purification via multiple chromatographic steps, which include specific heparin affinity, hydrophobic interaction, and ion exchange steps. A nanofiltration step is used as a precautionary viral removal step and glycine, sodium chloride, and sodium citrate are added as excipients. After filling into vials, the product is lyophilized. 

Product administration entails infusion after reconstitution with water for injection (WFI). Limited clinical trial results have been reported, mainly involving patients with hereditary AT deficiency in high-risk situations (surgery and pregnancy), with incidence of deep vein thrombosis. Common side effects include haemorrhage, headache, and nausea. Atryn's active ingredient is sourced from transgenic animals maintained by GTC biotherapeutics (Framingham, MA). Leo Pharma (Ballerup, Denmark) markets the product in Europe.  

(idursulfase) is a recombinant form of the human lysosomal enzyme iduronate-2-sulfatase, produced in an engineered transformed human cell line (HT-1080). The 525 amino acid enzyme displays a molecular weight of 76 kDa and harbors two disulfide bonds, eight N-linked glycosylation sites, and a formylglycine residue at position 59 that is necessary for biological activity. Elaprase, which is an orphan product, gained approval in the US in 2006 (and subsequently in the EU in January 2007). It is indicated for the long-term treatment of patients with Hunter syndrome (mucopolysaccharidosis II, MPS II). MPS II is a rare, X-linked recessive genetic lysosomal storage disease caused by a lack of functional lysosomal iduronate-2-sulfatase. This enzyme functions naturally in the body to cleave sulphate groups from two glycosaminoglycans (GAGs)—dermatan sulphate and heparin sulphate. GAGs are unbranched polysaccharides consisting of various uronic acids and hexosamine residues, which largely play structure-related roles in the extracellular space, particularly those associated with connective tissue. Lack of the enzyme results in GAG accumulation in most organs and tissues, resulting in organ or tissue dysfunction. Clinical manifestations usually become apparent within the first 12–36 months of life and are progressive, severe, and life threatening. 

Elaprase is manufactured through the initial culture of the human producer cell line, chosen for production due to its ability to attach complex, high mannose-type oligosaccharide side chains to the enzyme's polypeptide backbone. Specifically, the presence of mannose-6-phosphate (M6P) facilitates product binding to cell surface-M6P receptors, with subsequent cellular internalization and delivery to lysosomes—the exact target organelle.  

Product purification incorporates six chromatographic steps as well as two ultrafiltration and one viral filtration step. Formulation entails the addition of sodium chloride (to provide an isotonic solution for subsequent administration), sodium phosphate (a buffering agent) and polysorbate 20 (as a stabilizing agent and for protection against agitation-induced aggregation). The final product is filter sterilized and aseptically filled into presterile vials. Each vial contains 3 mL of a 2 mg/mL active ingredient solution. The recommended dosage regimen is once weekly intravenous infusion (after product dilution to 100 mL in saline) over a time course of 1–3 hours. 

Product safety and efficacy were established by a randomized, double blind, placebo-controlled trial containing 96 MPS II patients. Primary efficacy indications used were distance walked during a six-minute walk test and pulmonary function. The most common adverse events were infusion-related, including life-threatening anaphylactoid reactions. Elaprase is manufactured and marketed by Shire Human Genetic Therapies, Inc. (Danderyd, Sweden). 

 (insulin, human recombinant) is identical in amino acid sequence to native human insulin. It is produced by recombinant means in an engineered 

 K 12 strain, and gained approval for general medical use in 2006 both in the EU and US. Unlike formerly approved insulins, which are administered parenterally, Exubera is administered via the pulmonary route using a specially designed inhaler. It is indicated for the treatment of adults with diabetes mellitus, for the control of hyperglycemia. The inhaled insulin is absorbed more quickly than subcutaneously administered regular human insulin and has an onset of action that is similar to subcutaneously administered rapid-acting engineered insulin analogues, such as insulin lispro.  

After its absorption from the lung into the blood Exubera promotes a decrease in blood, glucose levels (and its other characteristic effects) in the normal manner, via interaction with the insulin receptor. 

The manufacture of Exubera consists of 16 steps. After initial fermentation, the 

 cells are harvested via centrifugation, with subsequent cell disruption and recovery of the insulin-containing fusion pro-product. Downstream processing includes a concentration step, a step to release free insulin from the pro-product and multiple chromatographic purification steps. Excipients then added include mannitol (a stabilizing and bulking agent) as well as glycine and sodium citrate (buffering and stabilizing agents). The product is spray dried, allowing its subsequent storage at room temperature. Product particles display a mean particle diameter of approximately 3 Âµm, which facilitates aerosol delivery to the deep lung. The finished product is filled into unit dose blisters containing 1 mg or 3 mg of active substance. These fit a manually operated, reusable inhaler device. 

Exubera has a duration of action comparable to subcutaneously injected regular human insulin and longer than rapid-acting insulin. It is administered immediately before food consumption and in regimens that include a longer acting insulin. When administered to healthy volunteers, the onset of (glucose lowering) activity was observed within 10 minutes with maximum effect noted after two hours and a total duration of about six hours. 

Clinical studies underpinning safety and efficacy were conducted using 2,500 adult patients with type 1 or type 2 diabetes, and the primary efficacy parameter monitored was invariably glycemic control. Like other insulin preparations, hypoglycemia represented the most commonly reported adverse event. Product administration was also found to cause a greater decline in lung function than controls and, as a consequence, the product is contraindicated in patients who smoke or who have poorly controlled or unstable lung disease. Exubera is distributed by Pfizer (New York City, NY) and licensed from Nektar (San Carlos, CA).  

 (human papillomavirus vaccine, types 6,11,16,18, recombinant), also marketed as 

 in the EU, is a multivalent vaccine preparation containing the purified recombinant major surface capsid (L1) protein of human papillomavirus (HPV) types 6,11,16, and 18, produced separately in engineered strains of S. 

. The product gained approval in 2006 in both the US and EU and is used to vaccinate girls and women (9–26 years of age) against cervical and vaginal cancer and genital warts caused by the HPV types listed. HPV infection represents the most common sexually transmitted disease worldwide, with some 50% of young women being infected within five years of becoming sexually active. Native papillomavirions are approximately 60 nm in diameter. These double-stranded DNA viruses display an icosahedral symmetry made by 72 pentamers of the L1 caspid protein. Some 40 different HPV strains preferentially infect the genitals and 13 of these are highly carcinogenic. They are believed to be the primary cause of cervical cancer, which is the second most common type of cancer causing deaths in women worldwide. Types 6,11,16, and 18 are the most commonly occurring HPVs and these are believed to cause the vast majority of cancers and genital wart growth.  

Gardasil manufacture requires  separate fermentation of four engineered S. 

 strains, each   housing the gene coding for the P1 capsid protein of one of the HPV strains listed. Following fermentation, the cells are recovered by microfiltration and the recombinant protein is released via homogenization. In each case, the recombinant capsid proteins spontaneously self-assemble into large virus-like particles (VLPs), rendering their subsequent purification— via size-based membrane filtration, ultrafiltration, and chromatography—relatively straightforward. The purified antigen (VLP) is then absorbed onto amorphous aluminium hydroxyphosphate sulphate, which acts as an adjuvant. Also added are sodium chloride, histidine, polysorbate 80, and sodium borate buffer as excipients. The product is presented as single use, 0.5 mL-doses in either vials or prefilled syringes with each dose containing 20 Âµg of type 6 L1 protein and 40 Âµg of each of the other L1 proteins. The administration schedule usually entails intramuscular injection of an initial dose, with subsequent doses delivered at months two and six. 

Clinical trials assessing product safety and efficacy involved more than 20,000 females and clearly showed a protective effect. The most common side effects noted were pyrexia (fever) and  reaction site injections. Gardasil is manufactured (and distributed in the US) by Merck (Whitehouse Station, NJ) and is marketed in the EU by Sanofi Pasteur (Lyon, France). It is also marketed in the EU as 

by Merck Sharp & Dohme (Hertfordshire, UK). 

 (ranibizumab) is a 48 kDa humanized MAb antigen-binding fragment (Fab fragment) consisting of a 214 amino acid light- chain fragment linked by an interchain disulfide bond to a 231 amino acid heavy-chain fragment. Produced in 

 the fragment binds vascular endothelial growth factor A (VEGF-A) with high affinity. It was first approved in the US in 2006, and subsequently in the EU (in January 2007). It is indicated for the treatment of neovascular (wet) age-related macular degeneration (wet AMD), a condition characterized by the formation of abnormal, leaky blood vessels under the macula (the central part of the retina). This condition can result in the lifting of the retina and hemorrhage in the subretinal space. As a progressive, degenerative disease, central vision is gradually blurred and the condition is most common in persons aged 75 years and older. 

VEGF-A, via binding to its endothelial cell surface receptors, is believed to be a major driving force for abnormal blood vessel formation. Binding of Lucentis to VEGF-A prevents interaction with the receptor, thereby reducing abnormal blood vessel growth. 

The manufacturing process of Lucentis involves initial fermentation of the 

 producer cell which carries cDNAs encoding variable light and variable heavy antibody chains fused to human constant light and constant heavy (CH1) domains. The light and heavy chains fold into their native conformation after their secretion into the periplasm. Soluble product is fully released from the 

 via initial homogenization and a subsequent heating step that precipitates cellular debris. High-resolution purification is achieved by a combination of four subsequent chromatographic steps, followed by the addition of trehalose, histidine, and polysorbate 20 as excipients.  

Lucentis is administered by direct intravitreal injection (0.5 mg active in a final volume of 0.05 mL) once monthly, and usually for an initial period of 3 months, with subsequent administration being dependent upon results from routine visual monitoring. 

The clinical efficacy and safety of Lucentis was assessed primarily in three randomized, double blind studies over a period of 24 months using a total of 1,323 patients (879 of which received Lucentis, with 444 untreated controls). The primary efficacy endpoint was either the proportion of patients who maintained vision (defined as losing fewer than 15 letters of visual acuity) compared with baseline or the mean change in visual acuity. The studies showed that Lucentis administration resulted in an increase in mean change in visual acuity as compared with continuous deterioration in the case of controls. The most common adverse reactions noted were conjunctival hemorrhage, eye pain, vitreous floaters, increased ocular pressure, and inflammation. Lucentis is manufactured by Genentech (San Francisco, CA) and is marketed in Europe by Novartis Europharm (Sussex, UK). 

(alglucosidase alfa) is a recombinant form of the human lysosomal enzyme acid α-glucosidase (GAA). The 109 kDa glycoprotein is produced in an engineered CHO cell line. It has designated orphan status in both the US and EU and was approved in both regions in 2006. Myozyme is indicated for the treatment of Pompe disease, a rare genetic condition characterized by a deficiency of lysosomal GAA activity. The condition is also known as glycogen storage disease type II, glycogenosis type II, or acid maltase deficiency. GAA functions to degrade lysosomal glycogen via the hydrolysis of both the α-1-4 and α-1-6 glycosidic bonds characteristic of the molecule. Pompe disease is characterized by accumulation of lysosomal glycogen stores in various cell and tissue types, particularly cardiac, skeletal, and respiratory muscle cells, leading to dysfunction of the effected cell types. 

The clinical presentation of the condition, which presumably is dependent on residual levels of catalytically active enzyme produced, ranges from a severe, rapidly fatal infant-onset form to a more slowly progressing late onset (juvenile or adult) form. 

Myozyme manufacture includes cell culture, product recovery, chromatographic purification, and formulation. It is presented in lyophilized form in glass vials containing 50 mg active, as well as mannitol, polysorbate 80, and sodium phosphate as excipients. The recommended dosage regimen entails intravenous infusion, once every two weeks over approximately four hours, at a typical dosage level of 20 mg/kg body weight. 

Product administration is followed by cellular uptake, likely through initial binding to cell surface mannose-6-phosphate receptors, with subsequent lysosomal delivery. Safety and efficacy were established primarily by two clinical trials involving a total of 39 Pompe disease patients ranging in age (at first treatment) from 1 month to 3.5 years. All were administered product and results were compared with historical data of (matched) untreated infantile-onset Pompe disease patients. Primary efficacy indicators included the proportion of patients who died or needed invasive ventilator support due to respiratory muscle failure. Both parameters were substantially improved with myozyme therapy. For example, historical control data indicated a mortality rate of 98% by 18 months of age compared with about 11% for the treatment group. The most common adverse effect noted is hypersensitivity reactions, including life-threatening anaphylactic reactions. Myozyme is manufactured and marketed by Genzyme (Cambridge, MA). 

(somatropin) is a recombinant form of human growth hormone (hGH) produced in an engineered 

. Like the native molecule, Omnitrope is an unglycosylated, 191 amino acid single-chain polypeptide with a  molecular mass of 22.1 kDa. It first gained approval within the EU in (April) 2006 as a similar biological medicinal product (a biosimiliar) and was subsequently approved in the US later that year. It is indicated for the treatment of children and adults with growth failure due to inadequate endogenous hGH secretion or also (in the EU) growth failure associated with chronic renal insufficiency, Turner syndrome, or Prader-Willi syndrome.  

Omnitrope binds the native hGH cell surface receptor, thereby triggering the characteristic direct and indirect growth-promoting and related effects of this hormone. 

The manufacture of Omnitrope is initiated by fermentation, during which the product accumulates intracellularly. Fermentation is followed by cell harvest and lysis, product recovery, and chromatographic purification. Glycine and phosphate buffer components are added as excipients and the final product is lyophilized in glass vials. Omnitrope is intended for daily subcutaneous administration with exact dosage levels being adjusted for individual patients. 

As a biosimiliar application the clinical evidence needed to support approval in the EU required demonstration of the product's similarity to a reference medicine (i.e., an hGH product already approved) in terms of quality, safety, and efficacy. The reference medicine chosen was Genotropin, an hGH product that first gained approval in Europe in 1988. Therefore, as required by EU biosimiliar legislation, the applicant was required to submit a full quality module and reduced clinical and nonclinical modules. Efficacy was demonstrated largely by a nine-month clinical study involving 89 children lacking hGH. Treatment with either Omnitrope or Genotropin yielded similar increases in height and speed of growth (equivalent to approximately 10.7 cm/year). This and a second trial involving 51 children who were administered product for a year also underpinned safety data. Side effects associated with Omnitrope (transient local skin reactions and in adults, mild edema, and muscle or joint pain) were also similar in type and severity to those associated with the reference medicine. 

The active ingredient is manufactured by Sandoz (Kundl, Austria), which also markets the product in both the EU and US.  

(parathyroid hormone) is a recombinant form of human parathyroid hormone produced in an engineered 

 cell line. The 9.4 kDa, 84 amino acid hormone is identical in sequence to the native human molecule and is devoid of disulfide bonds or a glycocomponent. It gained approval in the EU in 2006 and is indicated for the treatment of osteoporosis in postmenopausal women at high risk of fractures. 

Postmenopausal osteoporosis is characterized by low bone mass with resultant increase in bone fragility and hence increased risk of fracture. Parathyroid hormone functions as a primary regulator of calcium and phosphate metabolism in bones. It stimulates bone formation by osteoblasts, which display high affinity cell surface receptors for the hormone. It also increases intestinal absorption of calcium.  

The manufacture of Preotact involves initial (1100-L scale) fermentation of the producer 

 (cell line PAL 1000). The cells contain an expression plasmid (pJT42) housing a synthetic gene coding for human PTH fused to a 21 amino acid 

 outer membrane protein A signal peptide. As a result, the fusion product is secreted into the culture medium, with automatic removal of the leader sequence from the fusion product by an 

 signal peptidase. Following fermentation, the cells are removed from the product stream by a combination of centrifugation and filtration, followed by an ultrafiltration or diafiltration step. Purification includes five chromatographic steps, with addition of mannitol, sodium chloride, and citric acid as excipients, followed by lyophilization. The final product is presented as a dual chamber cartridge, with the second chamber containing the reconstitution solvent (WFI, containing metacresol as a preservative). The recommended administration schedule entails once daily abdominal subcutaneous injection of 100 Âµg active ingredient. 

The major clinical study underpinning product approval was an 18-month, double blind, placebo-controlled trial involving more than 2,500 post menopausal women. The trial demonstrated no significant difference in the incidence of non-vertebral clinical fractures, but demonstrated a 61% relative reduction of vertebral fracture (3.37% versus 1.32%) associated with product administration. Preotach treatment also increased the mineral density of the spine as well as hip bone. The most common side effects noted included increased serum and urine calcium levels (hypercalcemia and hypercalciuria) as well as nausea. The biologically active substance is manufactured by SynCo BioPartners (Amsterdam, the Netherlands) and it is marketed by Nycomed (Roskilde, Denmark). 

(natalizumab) is a recombinant, 149 kDa humanized IgG4  monoclonal antibody produced in engineered murine myeloma cells. In 2006 it was approved for resumed marketing in the US and approved for the first time in the EU. It is indicated as a monotherapy for the treatment of the relapsing forms of multiple sclerosis (MS) under certain conditions.  

Initial accelerated approval had been granted by the FDA in November 2004 on the basis of the substantial benefit recorded to MS sufferers as revealed by initial trials. At that stage, the most serious adverse events noted were acceptable infections and hypersensitivity reactions, but approval was contingent upon continuing the trials for an additional year. Marketing and its use in ongoing clinical trials was subsequently halted in February 2005 when the studies revealed two cases of  fatal progressive multifocal leukoencephalopathy (PML, a rare viral infection of the brain) in patients receiving Tysabri in conjunction with interferon therapy. The FDA allowed resumption of a clinical trial in February 2006 and the product subsequently gained approval under more limited conditions (e.g., it must be used as a monotherapy only, and it is made available under a restricted distribution program).   

The manufacture of Tysabri entails initial culture of the producer cells in a 15,000-L bioreactor, using media devoid of serum or animal-derived components. Cells are then removed from the antibody-containing media by membrane filtration and the product is purified by a combination of protein A affinity, anion exchange, and hydrophobic interaction chromatography. Additional (viral clearance) steps undertaken during downstream processing include low pH treatment and nanofiltration. The final product is formulated in the presence of sodium chloride, polysorbate 80, and sodium phosphate buffer components as excipients and is presented as a 15 mL-concentrate (20 mg active/mL) in glass vials.  

The recommended dosage regimen includes initial dilution of the concentrate in 100 mL saline, followed by once monthly administration via IV infusion over one hour. 

There were notable approvals in nonparenteral delivery systems and biosimiliars in 2006.

Biopharmecuticals: Approval Trends in 2006

The ability of an instantaneous microbial detection system (IMD-A) to monitor microbial populations in environmental air was evaluated. The IMD-A results were compared with results from conventional environmental air monitoring methods. The comparisons were carried out in controlled microbial barrier test chambers and in cleanroom environments. Additionally, microbial populations in environmental air in an unclassified environment were evaluated using the IMD-A and the all-gas impingement (AGI) method coupled with ScanRDI. In 1-m

 controlled-barrier test chamber studies the mean recoveries with the IMD-A were equal to or greater than the mean recoveries obtained with the Anderson air sampler at various concentrations. The mean microbial recoveries obtained using the AGI were higher, but in the same order of magnitude, as those recovered by IMD-A. In classified environments, microbial recoveries from the SAS air sampler were substantially lower than microbial counts detected by the IMD-A. There were reasonable correlations of microbial recoveries between the IMD-A and the SAS air sampler results in cleanroom environments. Mean microbial recoveries from environmental air in an unclassified environment were similar in the IMD-A and AGI methods coupled with ScanRDI analysis. These results suggest that the IMD-A has the potential to reliably and instantaneously evaluate microbial populations in environmental air to provide a valuable technique for biopharmaceutical manufacturing. 

Examining the microbial content of air is a key component of environmental monitoring in pharmaceutical cleanroom environments. Overall environmental air monitoring also includes evaluating the particulate content of the air. Typically, particulate content at 5.0 and 0.5 Âµm levels is measured using total particulate monitoring systems such as the Climet, PMS, Royco, Lighthouse, APC units, or similar systems.  

Evaluating microbial content in environmental air involves both active and passive air monitoring. Active microbial content in environmental air is typically evaluated using SAS, MAS, RCS, Mattson Garvin, Anderson air, liquid impinger, or SMA air sampling systems. Active air monitoring often involves the use of a device in which microorganisms from a known volume of air are captured on media plates, or alternatively, air is aspirated into a liquid and the microorganisms in the liquid are captured on a membrane filter and transferred to media plates to evaluate growth. Viable passive air is evaluated by the plate-count method using settling plates. Results from the microbial monitoring of the environmental air are typically not obtained until 3–5 days after sampling. 

That need to wait for several days to accommodate microbial growth before acquiring monitoring data has been a major limitation of conventional environmental monitoring methods. Over the past decade, several rapid culture- and nonculture-based methods have been developed to provide much faster turnaround for microbial data.

Table 1.  Statistical evaluation of 1-m3 microbial barrier test chamber data from the IMD-A and an Anderson air sampler-Bacillus atropheus (spores)			

One promising nonculture-based rapid method receiving increased attention in recent years involves the ScanRDI system (Chemunex, France).

 In this semiautomated system, the total number of viable organisms is determined by filtering samples through a membrane and labeling cells using a nonfluorescent substrate that diffuses across the cell membrane. This labeling differentiates between viable and dead cells based on the presence or absence of esterase activity and intact cell membranes. Only viable cells with membrane activity are able to cleave the dye and retain the fluorescent label. These viable microbial cells are quantified by scanning and counting using laser cytometry. Although the ScanRDI system offers the advantage of rapid evaluation of microbial populations, it is fairly specialized and does not allow for real-time detection of microbial populations. An ideal system for microbial air monitoring in pharmaceutical cleanroom environments would require little or no sample preparation or manipulation and would provide environmental microbial air monitoring data in real time. 

BioVigilant has developed a nonculture-based instantaneous microbial detection (IMD-A) system based on optical fluorescence sensor technology. The IMD-A system analyzes the quantity and size of biological particles in environmental air and simultaneously determines whether each particle is inert or biologic, all in real time.

 The IMD-A consists of three components: 1) an optical assembly to measure individual particle size, 2) a concurrent optical detector to detect a UV-laser-induced fluorescence signal from metabolites in microbial cells and spores, and 3) an algorithm for differentiating airborne microbes from inert dust particles. The optical assembly uses the Mie scattering detection scheme to accurately measure airborne particles with sizes ranging from 0.5–20 µm. Exploiting the Mie scattering detection scheme enables the use of ultraviolet (UV) light illumination to concurrently examine each particle for the presence of nicotinamide adenine dinucleotide (NADH) and riboflavin, which are necessary metabolic intermediates of living organisms, including bacteria and fungi. These metabolites are excited by the UV photon energy and subsequently by autofluoresce, which is detected by the sensor. Although the IMD-A cannot identify the genus or species of microbes, it can instantaneously determine the size of each airborne particle and whether it is biologic or inert. 

The overall objective of this study was to evaluate the IMD-A system in controlled and pharmaceutical cleanroom environments by comparing it with other microbial air monitoring methods. Specifically, we set out to do the following:  

Table 2.  Statistical evaluation of 1-m3 microbial barrier test chamber data from the IMD-A and an Anderson air sampler-Staphylococcus aureus (vegetative cells)			

1. Evaluate BioVigilant's instantaneous microbial detection (IMD-A) system in a microbial barrier test chamber (1 m

) in a microbial challenge side-by-side with conventional (Anderson air and air impingement) air-sampling methods 

2. Evaluate the IMD-A for use in cleanroom Class E (ISO 9), Class D (ISO 8, Class 100,000 at operational and static), Class C (ISO 7/8, Class 10,000 at static and 100,000 at operational), and A (Class 100, ISO 5) environments at the Bayer Berkeley site by running the IMD-A side-by-side with conventional (SAS) air sampling methods  

3. Compare the microbial recoveries in environmental air using the IMD-A with those obtained using ScanRDI.  

Note that classified areas mentioned above were monitored using areas that were in control, but not in use for manufacturing. 

 microbial barrier test chamber challenge study was performed at Nelson Laboratories (Salt Lake City, UT). In the first study, cultures of 

 spores were inoculated onto soybean casein digest agar (SCDA) and incubated for 2–7 days at 30–35 °C. The cultures were harvested in sterile water and the suspension was subjected to heat shock at 80–85 °C for 10 minutes to destroy vegetative cells. The titers of the spore suspensions were determined by serial dilution and plating onto SCDA, then adjusted to yield concentrations of 10

 barrier test chamber, bacteria were disseminated using a Chicago nebulizer. We performed experiments at five different spore concentrations, with tests repeated at least three times at each concentration. Microbial measurements of the environmental air in the barrier test chamber were done using the IMD-A (BioVigilant Systems, Inc., Tucson, AZ) and the Anderson air sampler (Anderson Instrument Company,  Fultonville, NY), a conventional device that relies on an impaction method to enumerate microorganisms in environmental air. SCDA plates from the Anderson air sampler were incubated for four days at 37 °C. In a second study, vegetative cells of 

 were used. Culture preparation was similar to that described above for 

 except that no heat shock treatment was performed. Experimental set-up was similar to that described above for 

 were tested at two different concentrations and the tests were repeated at least five times at each concentration. For both studies, data were normalized to account for the different flow rate between the Anderson air sampler and the IMD-A.  

 microbial barrier test chamber aerosol challenge study was performed at the US Army's Dugway Proving Ground (Dugway, UT). In this study, 

 spores were prepared similarly to the method described above for the 1-m

 microbial barrier test chamber challenge study. 

 barrier test chamber using a proprietary ultrasonic methodology that can specify the size and likely number of bacteria per aggregate to a high degree of accuracy. Nominal concentrations of 

 spores were dispersed over four runs: 30 cfu/L at 1.0 Âµm, 30 cfu/L at 2.5 Âµm, 300 cfu/L at 1.0 Âµm, and 300 cfu/L at 2.5 Âµm. Microbial measurements of the environmental air in the barrier test chamber were performed using the Anderson air sampler and using two IMD-A instruments, each placed at opposite ends of the chamber, and an all glass impinger (AGI), a glass bubble tube used to sample air. In the AGI method, a known volume of air is actively pumped through an impinger containing a liquid medium. SCDA plates from the Anderson air sampler were incubated for four days at 37 °C. To assess the AGI, liquid obtained was filtered onto a membrane to capture cells and the membrane was placed on a media plate and incubated to evaluate microbial growth. The membrane filters were transferred to SCDA media plates and incubated for four days at 37 °C. The results obtained from the Anderson air sampler, AGI, and the two IMD-A units were normalized to account for different flow rates in the three types of instruments.  

Table 3.  Statistical evaluation of 150-m3 microbial barrier test chamber data from the IMD-A, an Anderson air sampler, and AGI-Bacillus atropheus (spores)			

In another comparison test, microbial populations in cleanroom Class E (ISO 9), Class D (ISO 8, Class 100,000 at operational and static), Class C (ISO 7/8, Class 10,000 at static and 100,000 at operational), and Class A (Class 100, ISO 5) environments at the Bayer Berkeley Site were evaluated using the IMD-A, running it side-by-side with a conventional SAS air sampling method (Bioscience International, Rockville, MD). SAS air sampling uses sieve impaction: air is aspirated for a specific time period through a perforated cover plate. The air is then blown onto a solid agar media plate, where the particles are impacted. The plates are incubated at an appropriate temperature and time period and the colony forming units (cfus) are counted. Environmental air monitoring in the cleanrooms was performed over a period of eight weeks in areas where little or no manufacturing activities were taking place. Because of the newer device's lower flow rate, for Class A, C, D, and E, 35 liters of air were sampled using the IMD-A.  For Class A, C, and D, 1,000 liters of air were sampled using the SAS air sampler. For Class E, 200 liters of air were sampled using the SAS air sampler. Plates from the SAS air samplers were incubated for four days at 37 °C.  

Microbial populations in environmental air in an unclassified office space environment were evaluated using the IMD-A and AGI method coupled with ScanRDI analysis. One cubic foot of air was sampled with both the IMD-A and AGI. Then 100 mL of fluid (Fluid A) from the AGI was membrane filtered and processed for ScanRDI analysis in accordance with the manufacturer's instructions. We tested the samples in quadruplicate, with unused Fluid A serving as the negative control for the ScanRDI analysis.  

Figure 1A and Table 1 show summary results from the 

 microbial barrier test chamber aerosol challenge study. In general, with the exception of one concentration range tested (concentration 4), the mean biological counts from the IMD-A  were equivalent or greater than the mean 

 spore counts recovered by the Anderson air sampler. Although at the lower concentration ranges (concentrations 1, 2, and 3) the microbial counts from both instruments were more or less the same, at the higher concentration range (concentration 5) the biological counts recovered by the IMD-A were substantially higher than those recovered by the Anderson air sampler.  

Table 4.  Statistical evaluation of 150-m3 microbial barrier test chamber data from the IMD-A, an Anderson air sampler, and AGI-linear fit of instrument correlation			

 microbial barrier test chamber aerosol challenge study are shown in Figure 1B and Table 2. At both concentrations tested, the recovery of biological counts from the IMD-A were equivalent  to or greater than the mean 

 vegetative cell counts recovered by the Anderson air sampler. The mean biological counts recovered by the IMD-A were 50% and 25% higher than the microbial recoveries of 

Figure 2 and Table 3 show results from the 

 microbial barrier test chamber aerosol challenge study. At four of the concentrations tested, the biological counts recovered by the IMD-A instruments (IMD-A 1 and IMD-A 2) were equivalent. At all concentrations tested, the mean recoveries from the AGI method were higher than recoveries observed from the IMD-A instruments and the Anderson air sampler. The microbial recoveries from the Anderson air sampler varied when compared with the results from the IMD-A instruments and the AGI at each of the four concentrations tested. We observed a high degree of correlation for microbial recoveries between the two IMD-A instruments and between the IMD-A instruments and the AGI, as shown in Table 4. The degree of correlation between IMD-A and Anderson air sampler and between the AGI and Anderson air sampler were substantially lower. 

Microbial recoveries from environmental air in cleanroom environments Class E (ISO 9), Class D (ISO 8, Class 100,000 at operational and static), and Class C (ISO 7/8, Class 10,000 at static and 100,000 at operational) are shown in Figures 3, 4, and 5 respectively. In all three classified areas, the microbial recoveries from the SAS air sampler were lower than the biological counts recovered by the IMD-A. Correlations (r

) between the IMD-A and the SAS air sampler for Class E, Class, D, and Class C were 0.51, 0.42, and 0.32, respectively. Of the several replicated tests done in the Class A (ISO 5) environment, no biological counts were detected using either the IMD-A or the SAS air sampler (data not shown). 

We also assessed microbial recoveries from environmental air in an unclassified office space environment using the IMD-A and AGI methods coupled with ScanRDI (Figure 6). Mean microbial recoveries from the environmental air were nearly identical between IMD-A and AGI method coupled with ScanRDI.  

Results for the controlled microbial barrier test chamber studies demonstrate that the microbial recoveries obtained from the IMD-A units were comparable to or higher than the conventional culture based air sampling methods and indicate that the IMD-A has the potential to reliably evaluate microbial populations in environmental air. Mean microbial recoveries from the IMD-A were slightly lower than, but in the same order of magnitude as, those recovered by the AGI. On the other hand, microbial recoveries from the Anderson air sampling method were substantially lower than those recovered using the IMD-A or the AGI method.  

The strong correlation (Table 4) in microbial recoveries between the IMD-A and the AGI suggests that the IMD-A reliably detects microbial populations in environmental air. This is further supported by the fact that the mean microbial recoveries from environmental air were nearly identical between the IMD-A and the AGI method coupled with ScanRDI analysis (Figure 6). The slightly higher microbial recoveries attained with the AGI method in the microbial barrier test chamber can be explained by the fact that the impingement method is known to disperse microbial aggregates more effectively than other methods used to sample environmental air, while the IMD-A would potentially undercount microbial aggregates.  

However, since the IMD-A can simultaneously detect whether a particle is biological or not and determine its size, any potential microbial aggregates larger than individual cells can be readily flagged by the IMD-A. The lower microbial recoveries from the Anderson air samplers compared with the IMD-A or the AGI are not unexpected, since centrifugal, filtration, and impinger methods are thought to provide better recoveries and suggested to be more suitable methods for microbiological air monitoring than are impaction methods such as Anderson air samplers.

 created during impaction can limit the recovery of microorganisms on media plates. It is also well recognized that microorganisms in air may remain viable but lose the ability to form colony forming units, so that culture-based methods may underestimate microbial populations in environmental air.

In the classified cleanroom environments examined, microbial recoveries in environmental air as assessed by the IMD-A were substantially higher than those from the SAS air sampler, which recovers microorganisms by the impaction method. Reasonable correlation of microbial recoveries were observed between the IMD-A and the SAS air sampler in cleanroom environments (Figures 3, 4, and 5) Class E, Class, D, and Class C (r

 = 0.51, 0.42, and 0.32, respectively). Microbial recoveries from the IMD-A were approximately two orders of magnitude higher than those observed from the culture-based SAS method. Because of its slower flow rate, the IMD-A was operated for a longer time than the SAS air sampler. The higher recoveries may be partly explained by the fact that the IMD-A device could have captured microorganisms from activities in the cleanrooms subsequent to the end time of testing using SAS. 

Additionally, in natural environments, microorganisms can exist in viable, nonviable, or viable but nonculturable (VBNC) states, the latter of which is the predominant form.

 This is partly due to the fact that culture-based methods are very selective in their ability to recover microorganisms that often exist in symbiotic or syntrophic relationships in natural environments. There is little or no published information available about the VBNC phenomenon as it occurs  in controlled cleanroom environments. However, it is quite conceivable that cleanroom environments could have substantial levels of nonviable microorganisms, because  of the controlled environmental conditions in cleanroom environments that could be captured by the IMD-A, especially if the microorganisms recently lost their viability but still have substantial pools of metabolic cofactors such as NADH that persist and are detected by the IMD-A sensor.  

Results for the controlled microbial barrier test chamber, the ScanRDI study, and the cleanroom environment studies suggest that the IMD-A has the potential to provide a reliable evaluation of the microbial populations in environmental air and can detect variations in environmental microbial counts in cleanroom environments that are usually detected by conventional air sampling methods. While the microbial counts from the environmental air detected by the IMD-A in cleanroom environments were substantially higher than the conventional air sampling method, the ability of the IMD-A to simultaneously detect microorganisms in real time will prove the IMD-A to be a valuable analytical tool once its baseline for cleanroom environments is established.  

The IMD-A system can be valuable in monitoring environmental air in pharmaceutical manufacturing areas because of its ability to evaluate microbial quality of the environmental air in real time. There are several areas where we believe the IMD-A has potential immediate benefits:  

1) Investigations: If conventional environmental monitoring methods show unexpected elevated levels of microbiological contamination, or if media fill results in an aseptic area yield one or more positive vials, the IMD-A system would be a valuable tool to immediately assess the environment. Assuming the microbiological contaminants were still present 5–7 days later (which is not always the case), the IMD-A could help locate the contamination and ascertain its cause to facilitate corrective action. The IMD-A can also check the effectiveness of corrective actions during investigations in real time before operations are resumed.  

2) Reduction of shutdown time:  Currently, conventional environmental monitoring methods verify that an area has returned to its validated state after being shut down for repairs or renovations. Such environmental monitoring data determine when to restart the operation. In an aseptic environment, they are used to determine when to perform media fills. The IMD-A could supply information about the environment rapidly, reducing the risk of performing media fills before the environment has returned to normal. In combination with conventional environmental monitoring and media fill data, the IMD-A could provide additional information to collectively assess the risk of resuming operations faster than with competing methods. 

3) Training:  The IMD-A's ability to provide immediate feedback can be helpful to assess how an operator's movements in an aseptic environment could affect the product. This feedback can provide information about whether or not an operator is ready to be trained using conventional environmental monitoring data and media fills. 

Features

Evaluation of an Instantaneous Microbial Detection System in Controlled and Cleanroom Environments

 case shifted the landscape of patent licensing in an important way. As a result of the ruling, a patent licensee now can challenge the validity of a patent without having to breach the license first.  This ruling makes it easier for companies to protect themselves against patent infringement suits, and makes it harder for patent holders to enforce and license their patents.  

MedImmune had licensed a patent and a pending patent from Genentech related to the production of novel antibodies. The license required that MedImmune pay royalties to Genentech on valid, issued patents in exchange for permission to use those patents. Upon issuance of the patent for the application that had been pending, Genentech demanded royalty payments for the recently issued patent. MedImmune paid the royalties but said it was doing so under protest because it asserted that the patent was invalid and that its products did not infringe the new patent. MedImmune then filed an action under the Declaratory Judgment Act, asking the courts for "declaratory judgment," i.e., a ruling on the validity of the patent and whether it had been infringed.  

The lower courts dismissed the case for lack of jurisdiction, so the case was appealed and made its way to the Supreme Court. The Supreme Court held that MedImmune was legally entitled to file for declaratory judgment even while continuing to pay royalties.  

The Declaratory Judgment Act allows a party to petition the federal courts for a judgment about whether its rights have been violated. But such a declaratory judgment action can only be filed and ruled on by the court if there is a "case or controversy" between the parties.

  Until recently, this meant that to file a declaratory judgment action regarding patent validity or infringement, a party needed to have a "reasonable apprehension of suit" by the patent holder—i.e., a reasonable fear that the patent holder might sue for infringement. Under this definition, since a licensee cannot be sued for patent infringement unless it breaches the terms of the license, a licensee would only have a "case or controversy"—and thus, the right to seek a declaratory judgment about a patent's validity—if the licensee first terminated or breached the license, such as by refusing to pay royalties. By terminating or breaching the license, however, a company would open itself up to a lawsuit for infringement or breach of contract by the patent holder, thus leaving the licensee at considerable risk. 

 decision, the Supreme Court made an analogy to principles followed in cases against government action. In those cases, the courts do not require a plaintiff to transgress a law before challenging the constitutionality of the law in court, and a plaintiff's compliance with the law under protest does not eliminate the plaintiff's right to seek a declaratory judgment. Applying these principles to patent actions, the Supreme Court held that a patent holder does not have to infringe a patent before challenging the validity of the patent. The court did this by redefining "case or controversy," saying that a "case or controversy" exists when: (1) payment is demanded, such as pursuant to a license agreement or for infringement of a valid patent; and (2) the payment is made; but (3) the payment is involuntary or coerced. The Supreme Court found that the threat to MedImmune of treble damages and the loss of 80% of its business—which were the possible results of a patent infringement suit by Genentech—were coercive  enough to preserve MedImmune's right to file a declaratory judgment action.  

Genentech argued that MedImmune's suit was barred by the common law rule that a party to a contract cannot challenge the validity of the contract while continuing to reap its benefits. The Supreme Court noted that MedImmune was not repudiating the contract but was asserting that the proper interpretation of the contract did not prevent it from challenging the patents. Furthermore, the Supreme Court said that if the terms of the license agreement precluded the suit, Genentech would win its case on the merits rather than by obtaining a ruling that MedImmune did not have the right to file the suit in the first place.  

In the course of its analysis, the Supreme Court criticized the Federal Circuit test for declaratory judgment jurisdiction in patent cases. The Federal Circuit test required that there be both: (1) an explicit threat or other action by the patent holder that creates "a reasonable apprehension of imminent suit" and (2) current infringement activity or concrete steps taken towards infringement.

  According to the Supreme Court, its prior decisions allowing declaratory judgment jurisdiction when a licensee paid royalties under an injunction

  and when an appellate court affirmed non-infringement of the patent

  contradicted the Federal Circuit test. 

Thus, as a result of this Supreme Court ruling, patent holders now can be sued by licensees in good standing and as by prospective licensees who fear a patent infringement suit. This ruling provides a significant new protection to patent licensees and potential licensees who believe that they are being unfairly targeted by patent holders. 

 decision, the Federal Circuit further altered the patent landscape by holding that an offer to license a patent, in certain circumstances, may allow a declaratory judgment suit to determine patent validity and infringement—even if the patent holder promises not to sue. In 

STMicroelectronics (ST) owned several patents that it wished to license to SanDisk. During the course of several meetings, ST offered to license its patents and presented in-depth analyses of SanDisk's infringement. During these negotiations, ST representatives stated that ST had "no plan whatsoever to sue SanDisk." SanDisk, however, concerned that ST might in fact sue, filed a declaratory judgment suit against ST asking the court to rule on the validity of the patents and whether they had been infringed.  

 decision, the Federal Circuit abandoned its reasonable apprehension test and allowed SanDisk to bring its declaratory judgment suit. In the 

 decision, the Federal Circuit held that a "case or controversy" arises if a patent holder attempts to license its patent based on the activity or planned activity of a prospective licensee and if the prospective licensee contends that it does not require that license.  

In spite of these rulings, the specific actions of a patent holder that would give rise to the right to file a declaratory judgment lawsuit are not yet well defined by the courts. It is now clear, however, that a prospective licensee does not need to risk an infringement suit by engaging in the identified activity before seeking a declaratory judgment. The concurring opinion in 

 pointed out that this decision implies that practically any offer by a patent holder to license a patent opens the patent holder to a declaratory judgment lawsuit.   

In light of these recent decisions, patent holders must devise new strategies for patent enforcement and licensing. For instance, the concurring opinion in 

 suggested that holding negotiations under an existing confidentiality agreement may prevent a prospective licensee from being able to seek a declaratory judgment. It is still unclear, however, exactly how such a confidentiality agreement would affect a declaratory judgment suit, particularly since prospective licensees can now safely resist offers for confidential negotiations if they believe the patents to be invalid or not infringed, and licensees may file for declaratory judgments once the license is negotiated. Professionals representing licensors are now seeking to use various contract provisions in license agreements to impose adverse consequences if a licensee brings a declaratory judgment suit. 

is an associate in Perkins Coie LLP's licensing and technology group, Seattle, WA, 206.359.8069, 

 1. MedImmune, Inc. v. Genentech, Inc., 127 S. Ct. 764 (2007). 

 3. Altvater v. Freeman, 319 U.S. 359 (1943).   

In light of these recent decisions, patent holders must devise new strategies for patent enforcement and licensing.

BioPharm International-09-01-2007

From the Editor: Issues of Scale

Outsourcing: Robust Market Testing CROs' Ability to Deliver

FDA Leglislation Tackles Safety Issues

Biopharmecuticals: Approval Trends in 2006

Evaluation of an Instantaneous Microbial Detection System in Controlled and Cleanroom Environments

IP Briefs: The Shifting Landscape of Patent Licensing

Lifetime Studies for Membrane Reuse: Principles and Case Studies

Street Talk: Summer No Picnic for Investors

Final Word: In the Quest to Manage Capacity, Biology Trumps Stainless Steel