High-Fidelity PCR Reagents Continue to Propel DNA Research with Unparalleled Accuracy and Speed

New proprietary tracking dyes within high-fidelity DNA polymerase and qPCR master mixes facilitate easier, more accurate PCR set-up for researchers.  The Thermo Scientific Phusion Green High-Fidelity DNA Polymerase and Luminaris Color qPCR master mixes utilize proprietary tracking dyes to speed up the pipetting process. By providing a visual confirmation, the mixes are designed to vastly reduce the chance of human error, while also decreasing the number of procedural steps required. Most importantly, the dyes have been engineered not to interfere with the reaction or any downstream applications.

These new Phusion and Luminaris products form the latest additions to the renowned Thermo Scientific Phusion High-Fidelity DNA Polymerase family. First launched a decade ago, the Phusion High-Fidelity DNA Polymerase was the first DNA polymerase, developed using fusion protein technology to provide a combination of accuracy and speed previously unattainable using conventional enzymes.  Known for performance with all major PCR applications, the Phusion technology has played a central role in helping researchers accomplish an array of key scientific achievements, including the creation of the first functional synthetic genome.

“Phusion Polymerase technology has dramatically increased the productivity of labs running PCR,” says Margarita Leckiene, Director of Nucleic Acid Detection & Molecular Tools for Thermo Fisher Scientific. “Our philosophy is to simplify customer workflows further and increase the reproducibility of results. We are constantly working to further advance the accuracy and robustness of Phusion polymerase including the most recent innovation of special enzyme formulations that are optimized for NGS applications, offering equal amplification efficiency across entire genomes.”

This technology, coupled with Thermo Scientific PCR instruments and accessories such as theThermo Scientific Arktik Thermal Cycler and Thermo Scientific Piko Plate Illuminator, provides an efficient integrated solution that delivers

• DNA amplification with extreme accuracy
• Enhanced visual control
• Significantly shorter protocol times.

To recognize the impact of the original Phusion polymerase and a subsequent decade of innovation in this area, Thermo Fisher has launched Phusion Fest, an interactive campaign celebrating the importance of PCR in the laboratory. Phusion Fest encompasses a fun, scientific trivia game, special offers across a variety of molecular biology products and free samples of selected Phusion products.  More information on Phusion Polymerases and the Phusion Fest can be found by visitingwww.thermoscientific.com/phusion.

The Future of Drug Discovery: Two Required Supplements to Current Practices

A massive restructuring is currently taking place within pharmaceutical industry drug discovery sector.  Consequently, we suggest that phenotypic screening and drug repositioning will need to be increasingly utilized to uncover new therapeutics.

A convergence of forces in the pharmaceutical/biopharmaceutical industry continues to drive rapid rates of business restructuring in 2013. Although US regulatory policy issues, healthcare costs, globalization and other forces all contribute to this morphogenesis, the patent cliff that we are working through now has been a tremendous contributor as roughly $100 billion in sales have or will go off patent from 2010 to 2014. This massive loss in pharmaceutical companies’ top line is naturally flowing down to an array of cost-cutting measures in the industry, but drug discovery R&D is the true “canary in the coal mine.”

Since 2000, the pharmaceutical industry has eliminated about 300,000 jobs — as many people as currently work at the three largest drug makers (Pfizer, Merck and GSK) — combined.1 A significant portion of these are chemists, biologists and other scientists who participated in drug discovery. Large portions of R&D process have been outsourced to third-party, offshore contractors, raising criticism that true innovation, vital for drug discovery, is being significantly compromised.2 And although some have argued that drug discovery will increasingly shift into small to mid-sized biotechnology companies, and even academia, these sectors are also reeling from disruptions in the venture capital markets and pressures on the federal budget.

Phenotypic Screening

These disruptions and the restructuring of drug discovery across the industry have not significantly affected later stage, clinical programmes, as observed in rates of new drug application (NDA) fillings and new drug approvals, which have remained relatively stable (and even improved) in the last few years. It seems, however, quite likely that a day of reckoning is lurking on the horizon when opportunities for new drug candidates may begin to dry up. As a consequence, the industry has become very pragmatic about cost-effective strategies to drug discovery. Our experience indicates that strategic approaches such as phenotypic screening and drug repositioning will continue to be increasingly adopted in this new cost-effectiveness driven era of drug discovery.

The first of these strategies, phenotypic screening, can be viewed either as a departure or a complement to our existing standard paradigm for drug discovery — the target-based medicinal chemistry approach that runs central to all pharma R&D operations and has done since the 1980s and 1990s. The current approach, in a reductionist way of thinking, relies on our current understanding about biochemical pathways and their relationship to disease processes.

Literature Occurrences of Phenotypic Screening

Hypotheses are developed as to what enzyme or receptor should be modulated (inhibited or activated) to effect a positive outcome on a disease process. By contrast, phenotypic screening is, by its nature, not hypothesis-based, but instead is an empirical approach that relies upon observations of drug candidate activity in a system, such as an animal model of a disease process, independent of any initial hypothesis of why or how that candidate may be therapeutic towards the disease. One can make the argument, as have many pundits who have commented on the productivity gap in the pharmaceutical industry, that given the extreme complexity of biochemical pathways within intact higher organisms, that our hypotheses are too often wrong and the cost of testing them is too expensive leading to higher and higher R&D investments without concomitant productivity.3–5

The seminal publication by Swinney and Anthony showed, that despite an industry essentially focused on new drug discovery using the target-based approach, most first-in-class small molecule drugs were discovered by phenotypic screening.6 A possible criticism of phenotypic screening is that it will discover off-target effects that then need to be followed-up through additional hypothesis-based research. Based on our own mechanism of agnostic phenotypic screening of more than 200 drugs, we strongly believe that most (75–90%) of new biology uncovered by phenotypic screening is driven by on-target effects.7 An unbiased phenotypic screen is far more likely to uncover unexpected biology for a known mechanism than it is to discover new biology because of an off-target effect. Some companies such as Eli Lilly have been pursuing phenotypic drugs for close to a decade as a complement to mechanism/target-based drug discovery. Nevertheless, in our opinion, there is considerable opportunity to incorporate an increased balance of phenotypic to mechanistic screening in drug discovery. The barriers to increased phenotypic screening are partially the cultural familiarity with the mechanistic approach and in the case of cell-based assays the technical challenges of incorporating high-density data readouts into higher throughput assays.

Drug Repositioning

The second strategic element that is increasingly being adopted to increase cost-effective drug discovery is drug repositioning. It turns out that the famous words of Sir James Black: “The most fruitful basis for the discovery of a new drug is to start with an old drug” has a sound biological rationale that Black could not have fully appreciated at the time. From the point of view of molecular evolution, we now know that nature ‘recycles’ protein motifs again and again, and for this reason the chemical universe of biologically active compounds is characterized by dense spaces of hotspots with vast amounts of chemistry space that is biologically empty.8

Literature Occurences of Drug Repositioning

Although this finding is useful for medicinal chemists seeking to design new chemical entities, it also has a corollary, namely that compounds designed for one therapeutic area often have therapeutic benefit in other areas. In fact, it turns out that about 30% of approved drugs are labelled for indications other than the indication for which they were originally developed.9  Moreover a Thomson-Reuters Integrity database analysis shows that a drug in development for a single indication is the exception and that pursuit of multiple indications is the rule.10

Identifying new drug candidates from within the existing pharmacopeia (drug repositioning) has the well-recognized benefits of short cutting development, and thereby significantly reducing costs, by virtue of utilizing pre-existing preclinical and clinical drug development data. In most cases the first clinical studies for a new candidate can be in disease patients thereby obviating the time, expense and risk associated with Phase I studies. Although composition-of-matter patents typically are unavailable to provide exclusivity for repositioned candidates chosen from previously studied drugs, method-of-use patents, in many contexts, can be as rigorous (for compounds that have never had market approval). Also, with the lower costs to market it can be the case with many therapeutic product opportunities that the required commercialization thresholds may be met with the 5 years of data exclusivity available in the US and 10 years available in Europe through Trade-Related Aspects of Intellectual Property Rights (TRIPs) (again applies to compounds that have never had market approval).11 At a recent Washington DC drug repositioning conference there was audience unanimity that increasing the data exclusivity period was the single event most likely to enhance the drug repositioning field.

World’s Pharmaceutical Development Manufacturing Base Moving to India

The India Brand Equity Foundation (IBEF) has compiled its findings and released an overview of the current generics market in India. They predict that the Indian pharma market is now at the precipice of the next stage in its development, having seen manufacturing innovation and development technologies rise- thanks to the explosion in generics production.

During the last 3 years, exports of pharmaceuticals (largely generics) have grown at over 21.5% (CAGR) and now accounts for over $13bn in annual sales. Highlighting India’s dominance, nearly 40% of Abbreviated New Drug Applications (ANDA) received by the FDA in 2012 were from India, with a further 87 confirmed and another 25 already received between January and June 2013.

This huge growth in generics production has seen the country become a hotbed of manufacturing innovation – India has over 3000 DMFs registered with USFDA – which coupled with increased investments in R&D means India is now ready to challenge traditional big pharma and start producing more patented products. A natural evolution of the success of the generics market has been the rise in supergenerics across India where much R&D spend is currently being invested (e.g. Lincoln Pharma’s patent for NDDS).

With the world’s pharmaceutical development manufacturing base moving to India – there are 546 USFDA approved company sites (second only to the US), 23 companies holding 1100 authorizations with UK’s MHRA, and 166 companies with CEPs (Certificates of Suitability) from EDQM – coupled with the rise in supergenerics, the country’s next natural step is to use its world leading development expertise in the creation of new chemical entities.

Recognising this opportunity, the Government of India is putting in place supportive initiatives with the goal of cementing the country’s position as the ‘pharmacy of the world’ and creating a global innovation hub. With generics predicted to rise to 35% of global pharmaceutical market value by 2016 (some $400bn+), and with an annual growth rate of 27% amongst Indian generics exports (comparing very favourably with the global average of 10%) the Government and Pharmexcil are forecasting much of this revenue will be reinvested across the country in new research- leading to a steady pipeline of future drug targets.

In support of this, the Indian Government has committed to three schemes including a major multi-billion dollar initiative with 50% public funding through a public-private partnership (PPP) model to harness India’s innovation capability. In addition, the Government has made tax-breaks available to the pharmaceutical sector and a weighted tax deduction of 150% for any R&D expenditure incurred. Additionally, they have also introduced 19 dedicated Special Economic Zones to help stimulate pharma sector investment across the country.

Steps have also been taken to streamline procedures covering development of new drug molecules and clinical research- including two schemes ‘New Millennium Indian Technology Leadership Initiative’ and the ‘Drugs and Pharmaceuticals Research Programme’, which has been specially targeted at drugs and pharmaceutical research.

Already this year, India’s Dr. Reddy’s, Lupin Labs, Sun Pharma, Ranbaxy and Cipla have invested over $500million in R&D, which is allowing increased innovation in manufacturing processes  and will ultimately help to lower the cost of medicines production.

“India, termed as the Pharmacy of the World, has a basket of wide spectrum of generics that are second to none in terms of quality. The industry is on the track to expand its reach to newer markets, which makes it equally critical for the Indian pharmaceutical industry to keep its focus on quality, affordability and accessibility of medical solutions for the global pharma market. The country’s success in generics manufacturing is helping to keep our industry at the forefront of innovation and over the next few years we are lending our support to the R&D effort across the country so that we are leading in generics production and even developing new drugs out of India” said Mr Rajeev Kher, Additional Secretary, Department of Commerce, Ministry of Commerce and Industry, Government of India.

“Being a world leader in generics, India already has a huge presence in the highly regulated markets in terms of pharma exports. Almost two thirds of Indian generic exports are to the highly regulated markets (e.g. the US and Europe), which speaks volumes about the quality of Indian medicines. The Government of India is supporting Brand India Pharma campaign to reiterate that the Indian pharma market offers credible, affordable and sustainable healthcare solutions” said Dr P.V. Appaji, Director General, Pharmexcil.

About IBEF: India Brand Equity Foundation (IBEF) is a Trust established by the Department of Commerce, Ministry of Commerce and Industry, Government of India. IBEF’s primary objective is to promote and create international awareness of the Made in India label in markets overseas and to facilitate dissemination of knowledge of Indian products and services. Towards this objective, IBEF works closely with stakeholders across government and industry.

Foodborne Illness: Especially Dangerous for the Vulnerable

Older adults, pregnant woman and young children are among the most vulnerable to foodborne illnesses. People with compromised immune systems are also at risk.

If you've ever become sick after eating a food contaminated with disease-causing bacteria, it's not an experience you want to repeat.

But if you're part of what is called an "at-risk" or "vulnerable" population, a foodborne illness can be extremely dangerous. Symptoms—such as vomiting, diarrhea and fever—can intensify and the illness can become life-threatening.

Which populations are most at risk? According to Food and Drug Administration (FDA) epidemiologist Karl Klontz, M.D., M.P.H., they are the very young (under 1 year); older adults; the immune-compromised (those whose immune systems are less able to fight off harmful bacteria); and women who are pregnant.

For a particular type of bacteria known as Listeria monocytogenes—which causes a serious illness called listeriosis—the list is much the same, according to Vital Signs, a new report from the Centers for Disease Control and Prevention (CDC). The report, which summarizes data on the 1,651 listeriosis cases reported from 2009-2011, shows that older adults, pregnant women, newborns and persons with conditions that hinder the immune system are at a higher risk than others for listeriosis.

Combined, these vulnerable groups accounted for at least 90 percent of the listeriosis cases. CDC reported that 21 percent of the people with listeriosis died.

Immune System Plays a Role

What makes these populations more at risk? In many cases, the problem lies with the immune system, says Klontz. The immune system is the body's natural defense system against "foreign invasion" by pathogens (bacteria or viruses that can cause disease). In healthy people, a properly functioning immune system usually fights off harmful pathogens readily.

As we age, our immune functions and other barriers to infection start to wane, says Klontz. Our bodies less effectively fight off harmful bacteria. For example, the amount of acid in our stomachs, once a powerful barrier to pathogens, decreases. In addition, older people tend to take more medications for problems like heartburn or acid reflux, many of which further reduce the amount of stomach acid, further reducing this barrier against pathogens.

The same goes for people with compromised immune systems, such as those with HIV/AIDS, cancer, liver disease and diabetes. "Not only are their immune systems weakened by the disease," Klontz says, "but the side effects from certain treatments such as chemotherapy may make them weaker still."

On the opposite side of the age spectrum are children. Young children, in particular, are more at risk for foodborne illness because their immune systems are still developing.

As for pregnant women, "I wouldn't say that their immune systems are compromised so much as altered, serving a specific purpose—to enable the mother to co-exist with the fetus throughout the nine months of pregnancy. "Remember that half of the fetus' genes are not the mother's," Klontz says. The body has to work extra hard to avoid rejecting it. But that same alteration makes the body more susceptible to infection, he notes.

In addition, listeriosis in pregnant women can cause miscarriage, still birth, premature labor, and serious illness or death in newborns. Listeria monocytogenes, in particular, can cross the placenta (an organ which links the blood supply of mother to child) and infect the unborn baby.

Prevention is Key

Key to reducing the risk faced by these vulnerable populations is to prevent foodborne illnesses from occurring in the first place, says FDA microbiologist Mickey Parish, Ph.D. Prevention is at the heart of the FDA Food Safety Modernization Act (FSMA) signed into law in 2011.

FDA is working toward putting new measures in place to help keep contaminants out of the harvesting, processing and manufacturing of foods, Parish notes. By establishing safety and cleanliness requirements for farmers, food companies, and importers, FDA expects that implementation of FSMA will reduce the chances that pathogens such as Listeria, Salmonella, and E.coli will reach those most at risk.

There are steps that the people particularly vulnerable to the dangers of foodborne illnesses can take to reduce that risk, says Klontz. These include:

• Avoid eating raw animal products, which include unpasteurized milk (and cheeses made from unpasteurized milk), uncooked or lightly cooked eggs, and raw fish and meat dishes such as sushi or steak tartare.
• Wash fruits and vegetables before eating, especially foods with rinds, such as cantaloupes and other melons. Avoid eating raw sprouts.
• Make sure counters and other food preparation surfaces are adequately cleaned.
• Avoid hot dogs and other deli-style meats unless they are reheated to steaming temperatures. Also avoid deli-prepared salads, such as chicken or seafood salad.
• Keep your refrigerator at 40 degrees F or lower, and your freezer at 0 degrees F or lower.

If you're eating out, notes Klontz, especially if you're in an at-risk group, it's helpful to ask what ingredients are in a prepared dish. Are any raw or uncooked? Is the salad dressing or sauce made with unpasteurized milk or eggs? Does it include any raw animal products?

This article appears on FDA's Consumer Updates page, which features the latest on all FDA-regulated products.

Cancer Research UK installs Flexicon FP50

Cancer Research UK Biotherapeutics Development Unit (BDU) has installed a Flexicon FP50 tabletop filling and stoppering machine from Watson Marlow Pumps Group

A Flexicon FP50 tabletop filling and stoppering machine from Watson-Marlow Pumps Group has automated a previously manual process at a Cancer Research UK facility in Hertfordshire, UK. The FP50 is delivering increased speed and reliability into this critical operation, as well as enhanced vial filling accuracy.

The Cancer Research UK Biotherapeutics Development Unit (BDU) was built in 2010 at South Mimms. It is a modern, MHRA licensed, fully cGMP-compliant, 2000m²facility that is engaged in the process development and GMP production of Investigational Medicinal Products (IMPs) for Phase I clinical trials sponsored by Cancer Research UK.

Recent challenges at the BDU included the introduction of an automated filling and stoppering procedure, which was previously performed manually using a dosing pump. This method was not only labour-intensive and slowed down throughput, but was also unsatisfactory for such a leading edge facility. A reliable automated vial filling process was therefore required for the delivery of future BDU projects.

‘To help deliver this we set out to source a machine capable of rapid, repeatable filling but without any compromise to process quality and control,’ said Tim Hillyer, Senior Scientific Officer at the BDU.

The flexibility offered by the FP50 was ideal for our purposes

The Flexicon FP50 tabletop filling and stoppering machine from Flexicon Liquid Filling, part of the Watson-Marlow Pumps Group, is capable of filling up to 25 vials a minute (up to 100ml capacity). It also offers quick and easy changeover between batches.

‘As a multi-product facility producing small batch size, high-value IMPs, the flexibility offered by the FP50 was ideal for our purposes,’ said Hillyer. ‘Our most recent batches involved not more than a few litres in quantity so having a reliable method for product filling and stoppering enables us to bring new drugs into the clinic in a controlled and reproducible manner.’

Hillyer said the FP50 is quicker than the old manual process by a ratio of several factors. It is also easy to adapt for different vial and stopper sizes. An adjustable walking beam transports vials from the feeding turntable to the different working positions (filling needle and stopper plug) and only two parts need to be changed to cater for the entire range of vials and stopper sizes used by the BDU. The pump is also accurate, allowing the BDU to control the dose volume going into each vial.

The FP50 unit is fully contained within a six-glove isolator allowing the unit to gas the equipment prior to each product fill using hydrogen peroxide vapour. This containment twinned with the speed and efficiency of the FP50 greatly reduces the chances of batch contamination.

The Flexicon FP50 is a universal and aseptic tabletop filling system with integrated full or partial stoppering of rubber stoppers for use in pharmaceutical R&D departments and biopharma facilities. All materials and surfaces are designed to meet cGMP standards for aseptic filling, thus providing a ready-to-use validated filling system to carry out clinical trials and small batch production. The filling accuracy of the peristaltic filling system is better than ±1%.

The operator interface is an easy-to-clean touch screen and keypad. The panel is mounted on a separate control box, remote from the filling unit. This allows the control panel to be placed outside the LAF bench or isolator. It is possible to store up to 20 sets of filling parameters as complete working programmes.

Novartis Japan Achieves Primary Endpoint In HER2 Positive Advanced Breast Cancer Phase III Afinitor Trials

Everolimus

Novartis announced it achieved its primary endpoint of significantly extending progression-free survival with Afinitor (everolimus) in Phase III trials of patients with HER2 positive advanced breast cancer.

Everolimus (RAD-001) is the 40-O-(2-hydroxyethyl) derivative of sirolimus and works similarly to sirolimus as an inhibitor of mammalian target of rapamycin (mTOR).

It is currently used as an immunosuppressant to prevent rejection of organ transplants and treatment of renal cell cancer and other tumours. Much research has also been conducted on everolimus and other mTOR inhibitors for use in a number of cancers.

It is marketed by Novartis under the tradenames Zortress (USA) and Certican (Europe and other countries) in transplantation medicine, and Afinitor in oncology.

Sanofi Updates Lantus Label in EU, ORIGIN Results on Lantus® Cardiovascular Safety Integrated Into European Union Product Label

Lantus® (insulin glargine)

Sanofi  announced today that the Committee for Medicinal Products for Human Use (CHMP) of the European Medicines Agency (EMA) has issued a positive opinion for inclusion in the Lantus® (insulin glargine) product label of safety and efficacy data from the insulin glargine cardiovascular (CV) outcomes trial ORIGIN (Outcome Reduction with Initial Glargine INtervention). The revised label is evidence of Sanofi’s ongoing commitment to further assert the well-known safety and efficacy profile of insulin glargine, the most-studied basal insulin. The indication for the use of Lantus® remains unchanged.

MECHANISM OF ACTION
Insulin glargine (lantus) mechanism of action.

LANTUS (insulin glargine rdna origin injection) consists of insulin glargine dissolved in a clear aqueous fluid. Each milliliter of LANTUS (insulin glargine rdna origin injection) contains 100 IU (3.6378 mg) insulin glargine.

LANTUS® is a sterile solution of insulin glargine for use as an injection. Insulin glargine is a recombinant human insulin analog that is a long-acting (up to 24-hour duration of action), parenteral blood-glucose-lowering agent.,  LANTUS (insulin glargine rdna origin injection) is produced by recombinant DNA technology utilizing a non-pathogenic laboratory strain of Escherichia coli (K12) as the production organism. Insulin glargine differs from human insulin in that the amino acidasparagine at position A21 is replaced by glycine and two arginines are added to the C-terminus of the B-chain. Chemically, it is 21A- Gly-30Ba-L-Arg-30Bb-L-Arg-human insulin and has the empirical formula C267H404N72O78S6 and a molecular weight of 6063.

An Improved Manufacturing Process for the Antimalaria Drug - Artemether

Artemisinin and its derivatives, such as Artemether, are highly sensitive compounds, which require careful optimized production processes for their manufacture. Due to robustness issues, the manufacturing procedure of the reduction of Artemisinin with potassium borohydride to dihydroartemisinin was reinvestigated. The most important factor for obtaining optimal yields is to ensure low levels of contamination of potassium hydroxide in potassium borohydride. Application of a lower reaction temperature, fast addition rate of potassium borohydride, and careful control of the pH during the quench with acid are further important parameters in guaranteeing a robust process. In the redesign of the conversion of dihydroartemisinin to artemether, the yield was increased, and dichloromethane was replaced by the ecologically friendlier methyl acetate. A robust manufacturing process forartemether is now at hand, allowing the production of this important medicine reliably and in good quality and yield.

Process Validation Guidance: A Bad Fit for Aseptic Processing?

When he first assessed FDA’s draft Process Validation guidance a few years ago, consultant James Agalloco, president of Agalloco & Associates, saw the usefulness of the guidance for validating pharma production processes and products. “The life-cycle model will result in development and validation exercises that provide relevant and meaningful information,” he wrote. “The link between the process parameters that influence the critical quality attributes will serve the industry well. The use of statistical methods will add a rigor to the validation efforts that has been sorely lacking” [1].

However, he expressed serious reservations (and shared them with FDA) about whether the PV guidance could easily be applied to processes and systems “less clearly related to end-product quality attributes.” This includes sterilization and aseptic processes.

“There are simply too many independent—and interrelated—variables in aseptic processing, and the most meaningful one of all lacks metrics of any type . . . Aseptic processing performed by human operators is devoid of any measurable variable that could be used to predict the outcome.” He concluded: “The statistical component of the guidance really doesn't work with respect to linking any process parameters directly to performance.”

Now that the guidance is official, we checked in with Agalloco to see if his views have softened. Far from it, it turns out. The guidance is a “terrible fit” for the validation of aseptic processes, he maintains.

“I've seen more confusion than clarity,” Agalloco says. “I've seen no indication in the final guidance or anywhere else that suggests things should be changed to accommodate sterility within the new guidance.”

Agalloco teaches regular courses on validation of aseptic processes, but has not changed them to reflect the new guidance. “There's nothing you can change in the practice of either sterilization or aseptic processing that is of any value to fit the guidance,” he says. “I see no need to adapt, because the adaptation will gain nothing of real value.”

The new guidance reinforces the need for science and process simulation testing—isn’t that a good thing? In general, yes, he says. But, “for aseptic processing its about set-up and interventions, and what the operator does in relation to them. Humans are awful subjects for DoE, QbD and all of the scientific buzz we hear about in relation to the guidance.”

Will the PV guidance dramatically change the way manufacturers approach the validation of sterilization processes—filtration sterilization, for example?

“Not in the least,” Agalloco says. “There's no suggestion that anything can or should be changed to match the guidance. Even FDA (Grace McNally, at PDA in San Antonio in April of this year) has stated that the ‘guidance doesn't specifically apply to sterilization.’ Making changes to fit these processes to the guidance is done at one's peril.”

The PV guidance recommends activities in three stages (process design, process qualification, and continued process verification), which depend on process characterization studies (with Key Process Input Variables, Design Space, etc.). Are most manufacturers able to adequately characterize their aseptic processes and sources of variability?

Says Agalloco: “Not at all. As the article [1] states, there's some possibility with sterilization as it relates to Stage 1, but that's about all that fits. Stage 2 & 3 lack adequate metrics to develop the confidence from what we can routinely measure in the process for sterilization so it's not a good fit there. Aseptic processing is just a horrible fit all around.”

 

We also consulted with Sartorius Stedim’s Maik Jornitz, current chair of the board at PDA, about whether the guidance will make a difference regarding the validation of filtration processes, one of his areas of expertise. “I do not think that the new Process Validation Guidance will change the process validation needs and activities surrounding sterilizing grade filtration,” he says. “The 2004 Aseptic Processing Guidance makes it very clear what is expected by regulators in regard to sterilizing grade filter validation, which is still very valid and unchanged by the new Process Validation Guidance.”

Will this guidance help filter end users to better understand which filter process input variables are "critical to quality"? “I do not know whether it will help, but it will raise the awareness and emphasis that there are critical variables within the filtration process,” Jornitz says. “The 2004 Aseptic Guideline touches upon and PDA Technical Report #26 describes in detail these critical process parameters, if the awareness is not already there.”

Jornitz, a frequent writer, speaker, and lecturer, has long preached a “science-based” approach to filter validation, and so believes the PV guidance will support this trend but is not necessarily a game-changer: “I believe everybody in the industry is working in accordance to a science-based approach,” he adds. “Therefore, guidances are only there to reiterate or support what should and is commonly done—or so I hope . . . I do not think that the Process Validation Guidance states something new here.”