Chris Spivey is director of Industry Relations and Strategic Partnership, MJH Life Sciences.

Chris Spivey

Cytiva's Eric Corti discusses the challenges with designing a new single-use mixing system that ensures leak-free fluid handling operations.

Meeting Upstream Bioprocessing Challenges with Innovative Engineering Design (INTERPHEX 2024)

With the recent launch of Cytiva’s new single-use magnetic mixer system, the company took to 

® at the show, Eric Corti, global product manager for the Mixer Portfolio at Cytiva, discussed the key characteristics, challenges, and future of design of such equipment.

“This is a large-scale mixer that is primarily targeting media and buffer preparation applications. It's solving some really unique challenges in that space, and that's really driven by the consumable,” Corti stated. He explained that, in talking with customers, the company came to understand that one of the main problems plaguing the industry for large-scale media and buffer preparation are leaks.

To address this problem, the company aimed to design consumables that could effectively protect against leaks. The consumables were thus designed with protective features, such as a carrying case that wraps around the outside of the film and a covering for the impeller for protection on the inside. The company’s work didn’t stop at the design alone, Corti remarked, but rather the company went a step further to simplify the process as well.

“We understand that, during installation, large-scale consumables can often be tricky to install, and if they're done incorrectly, this can compound and lead to leaks, so we wanted to design something that was really simple to install, made it ergonomic and intuitive for operators to place the consumable inside of the hardware, and overall just make that experience [much] simpler and take [up] [much] less time. [This] can cut down on these leaks occurring in the first place, and can save customers a [significant amount] of money and a lot of pain and trouble that comes when those leaks do occur,” Corti emphasized.

Corti also pointed out that the new mixing system does not require any special configuration of the facility. “The mixer here has been deliberately designed to have a footprint that is going to fit through double doors that are standard for most clean rooms, as well as fitting within typical cleanroom ceiling heights. It’s going to cut down [much] of the costs … typically associated with modifying facilities just to get large-scale equipment in the door,” he explained.

Furthermore, Corti predicts that the future of single-use mixing systems design is leading more toward higher quality and increased quality assurance. Building on those protective capabilities in the design, Corti stated that many end users are asking for things such as point-of-view sleep testing and the ability to confirm even further that, once the consumables are installed in the system, there is a guarantee that there will be no leaks.

INTERPHEX 2024 was held in New York City on April 16–18, 2024.

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Global product manager at Cytiva, Eric Corti, discusses the challenges with designing a new single-use mixing system that ensures leak-free fluid handling operations at INTERPHEX 2024.

PharmTech Interview with Dennis Powers from G-CON at INTERPHEX 2024

As biologics continue to experience growth in the biopharma industry, access to appropriate cleanroom facilities that can provide a suitable and sterile environment is increasingly in need. In an interview at 

 spoke with Dennis Powers, senior vice-president, Product and Strategy, G-CON Manufacturing to discuss podular cleanroom solutions.

Primarily, G-CON had developed a standard POD cleanroom platform for the cell and gene therapy field, where a smaller footprint can aid in improving efficiencies. “The standard pods are well suited for cell therapy applications, small footprint aseptic, filling applications, [and] process development labs,” Powers said. Over the years, the company has developed a number of prefabricated designs that can be applied to different applications and customized depending on the manufacturers’ needs, he added.

With regards to the market for prefabricated podular/modular cleanroom solutions, Powers noted that there is definitely a rise in the number of companies in the field now. “We’re certainly seeing the industry starting to recognize the value in prefabricated modular construction,” he stated.

Additionally, modularization is not only changing facilities but is also being implemented into processes as well, Powers remarked. “So, as processes become more modular and more configurable [with a] smaller footprint, they fit very well into a prefabricated modular cleanroom infrastructure,” he said.

However, given the fact that the biopharma industry is more conservative than others, adoption of new technologies and approaches can be slower. “So, you have to not only have a great product, but you also have to educate the industry on that product, the value that product brings, and convince them why it's better than what they've been using traditionally,” Powers emphasized.

While at the show, the company launched the floorlessPOD, which is a freestanding structure that can be installed directly to the host facility floor, Powers explained. “[The floorlessPOD] allows us to eliminate the need for ramps platform steps to get up into the cleanroom, [and] it allows for direct integration into sub-slab utilities floor drains,” he said.

At INTERPHEX 2024, Pharmaceutical Technology® chatted with Dennis Powers from G-CON Manufacturing about podular cleanroom options and the benefits they offer for manufacturers.

Looking at the Benefits of Podular Cleanroom Solutions (INTERPHEX 2024)

Vial and injection syringe with DNA chromosome. Health care research, gene editing, molecular biology science, cell genetic analysis, medical DNA genetic engineering technology science background. 3D | Image Credit: ©Corona Borealis - stock.adobe.com

Sudden demand for large-scale manufacturing of messenger RNA (mRNA) drug substances during the COVID-19 pandemic has few parallels in the history of pharmaceuticals. Some might posit penicillin production during World War II as an analog, or perhaps early insulin availability. Pandemics and epidemics such as smallpox, typhus, cholera, influenza, HIV/AIDS, or the bubonic plague, all relied more on infection control for resolution than on manufacturing capacity. New therapeutic uses for mRNA are now ramping up good manufacturing practice (GMP) demand, especially for clinical trial supply. Moderna collaborated with CARsgen Therapeutics to combine an mRNA therapeutic cancer vaccine with a solid tumor chimeric antigen receptor (CAR)-T cell therapy, aiming to treat multiple cancers (1). Pfizer partnered with Beam therapeutics to co-develop mRNA and base editing approaches (2), while BioNTech and DualityBio have a partnership to develop differentiated antibody-drug conjugates, for solid tumors (3). In terms of upstream manufacturing for novel mRNA drug products, this technical complexity forms a tidal wave of question marks.

A prominent paper (4) examined reducing complexity by evaluating what common process could be standardized through extrapolations of clinical trial complexity, based on four published papers Phase I-II dose ranging studies. While not the amounts of product required during the COVID-19 pandemic, clinical trial doses quickly mounted up in terms of both volume and also manufacturing challenges related to techniques pursued, but also formulation delivery requirements. The authors state, “from the four studies, the average mRNA usage was 5500 μg and for the 75 trials in 2021, 412 500 μg of RNA would need to be formulated, purified, sterile filtered, and fill-finished. At concentrations of 100 to 200 μg of mRNA per mL, this would amount to 2063 to 4125 mL of drug product” (4). As an interesting foray into possible future manufacturing, the authors went on to contrast traditional mRNA with a self-amplifying method. Often, this is cited as potentially a 100 to 1 decrease in required finished product due to higher potency performances, but this study chose a picturesque perspective, saying, “assuming the entire population consists of 7.7 billion people, and each person receives two doses, a total of 15.4 billion doses or 4.5 million L of the vaccine is required. In perspective, 4.5 million L would fill two Olympic-sized swimming pools (2500 m3), whereas saRNA [self-amplifying RNA] vaccines, which we assume are 100-fold more potent, would only require 0.02 Olympic-sized swimming pools. Assuming that production costs, which account for materials, RNA, and LNP [lipid nanoparticles] production, scale linearly, vaccinating the entire population with Pfizer/BioNTech’s mRNA vaccine would cost approximately 150 billion USD. In contrast, with saRNA, it would only cost approximately 1.5 billion USD” (4).

Commercial-scale mRNA is produced through chemical, recombinant, or enzyme-based methods. For chains under 100 nucleotides, the standard chemical technique “utilizes phosphoramidite chemistry and solid-phase support promoting chain elongation from the 3′ end to the 5′ end” (2). It is amenable to automation, as is recombinant production through host cells such as 

), which has some cost advantages as it can utilize legacy protein manufacturing lines, already in GMP compliance. Enzymatic processes are newer to arrive and have tradeoffs still being worked through. A study conducted by the Duke Human Vaccine Institute explored creating standardized robust manufacturing processing steps, saying they confronted the lack of established quality standards by, in part, surveying vendor operations “through questionnaire and/or audit relating to the vendors’ quality systems and operations as they pertained to manufacturing and/or testing of the materials” (5). The study adopted the Weismann–Karikó method of production, the upstream process requiring three enzymatic steps of plasmid linearization, mRNA transcription, and DNA template digestion. In the starting out phase, fundamentals the authors emphasized were that “for optimal 

 transcription yields, it is critical to start with high quality template DNA. Some key quality attributes that are required before use include the presence of only a single band of linearized DNA, a supercoiled ratio of at least 70%, and a sequencing-verified correct poly(A) tail length” (5). They also noted that “endotoxin control was the primary raw material risk due to its variability in raw material lots and the difficulty of its removal in subsequent downstream purification steps … reaction agitation, temperature, incubation time, and reactant concentrations were also carefully examined for each reaction step in order to yield a robust upstream platform process across multiple mRNA constructs” (5).

Chris Spivey is director, Industry Relations and Strategic Partnerships

Articles

A tidal wave of questions floats the need for more upstream automation.

The Making of mRNA

®, Susan Schniepp, distinguished fellow, Regulatory Compliance Associates, and co-chair of board of directors, Parenteral Drug Association, expands on the importance of maintaining a robust quality management system (QMS) in bio/pharmaceutical manufacturing. Schniepp hosts an interactive session on the impact of QMS on quality maturity at INTERPHEX, held in New York City on April 16–18, 2024.

For advanced therapy medicinal products (ATMPs) in particular, Schniepp emphasizes how fast-moving this sector is. “The regulations don't keep up with the ATMPs. That technology, and their way of thinking, is turning over quicker than the regulations,” she says.

“The changes in the regulations that are going to come are going to be around quality culture and maintaining a robust quality management system,” she adds. Ensuring documentation and keeping equipment calibrated are important practices to apply to these new fast-moving ATMPs, she states. Schniepp does not necessarily expect to see many changes in the regulations around ATMP development and manufacturing but thinks that there will likely be more guidance documents issued in the future, with one of FDA’s focuses being its quality management maturity model.

“There are some regulations out there that call out quality culture. In particular, the World Health Organization has one on data integrity. It has a definition and standard[s] on what quality culture is,” Schniepp says. She points out that a new aspect of her presentation at INTERPHEX this year is its interactive component, in which she sets up a scenario involving an audit where an incident occurs. She gives the audience three potential responses to discuss, but rather than simply asking them which response do they pick or which response is correct, she instead asks what does the chosen response say about that person or that company’s QMS and the maturity of that system?

Susan Schniepp, distinguished fellow, Regulatory Compliance Associates, demonstrates QMS impact on quality maturity at INTERPHEX.

Susan Schniepp of Regulatory Compliance Associates Explores the Impact of QMS on Quality Maturity (INTERPHEX 2024)

Susan Schniepp, distinguished fellow, Regulatory Compliance Associates, and co-chair of board of directors, Parenteral Drug Association, demonstrates QMS impact on quality maturity at INTERPHEX.

® speaks to Bill Whitford, Strategic Solutions leader at Arcadis, on the role and impact of 3D bioprinting for biologics. Whitford is a presenter on the topic of 3D bioprinting at INTERPHEX, held in New York City on April 16–18.

Whitford notes that an important aspect of 3D bioprinting is that it is one approach to 3D cell culture, which he observes is taking over cell culture in many categories. “3D culture provides many unique capabilities it's more biomimetic. Actually, there's nothing natural about growing cells in monolayer on a styrene plate, so there are many 3D culture technologies that employ many different production methods that support many categories of application and individual products,” Whitford states.

On the workflow in a 3D bioprinting facility, Whitford remarks that many complementary and supportive synergistic technologies exist that support each other. “For example,” he explains, “machine vision now is really simple. Ordering 3D printing and the ability to both direct and inspect a nascent print job using artificial intelligence (AI) … they feed on each other. The availability of bioprinters and what we call bio ‘inks’ is, in a commercial sense, what's really driving the final applications.”

“Best practices are being developed for bioprinting,” Whitford also notes, pointing out that there are many different ways of affecting bioprinting, not just with respect to the instrumentation, but with respect to the actual printing. “There is laser-effected printing; there's extrusion-effected printing. There are many different ways of getting the bio ink into the three-dimensional structure, and they all have their advantages. They all provide either a tremendous accuracy and precision or volume.”

In the interview, Whitford also discusses the future of 3D bioprinting in the cell and gene therapy sector. “The ability now—through site-directed CRISPR [clustered regularly interspaced short palindromic repeats]-type engineering—to drive the evolution of a cell towards exactly what phenotype is required is supporting the ability to … develop the bio ink … required to get the initial print job accomplished.”

At INTERPHEX, Bill Whitford, Strategic Solutions leader at Arcadis, discusses the progress made in 3D bioprinting toward commercial biologics production.

Bill Whitford of Arcadis Discusses 3D Printing for Biologics (INTERPHEX 2024)

Bottles for injection with white powder on a color background | Mr.Ilkin

There are two main threats posed by extractables and leachables (E&L), contamination from unwanted impurities such as nitrosamine or chromium causing toxicity issues and diminution of drug efficacy or potency by destabilizing or perturbing the API. From a system-wide perspective, the E&L threat posed by manufacturing equipment is small, versus lifetime container closure and material packaging of drug products. The effects of the former are comparatively sudden and obvious (and hence easier to spot), versus the slow chemical transformation that might occur in the latter. In this sense, leachables are a subset of their parent extractables.

Recalls of nitrosamine-contaminated drugs “underscore the need for robust analytical methods and risk-based approaches for E&L and nitrosamines testing. Additionally, drug product manufacturers must ensure that toxic elemental impurities are under a state of control in their products” (1). There are many courses of instruction one can take to learn how to implement good monitoring of E&L. In line with wider industry trends, risk-based approaches to E&L testing for small molecules, especially, are becoming more commonly implemented, and the salient issue of nitrosamine contamination has certainly accelerated that trend.

Plasticizers, stabilizers, lubricants, or colorants are well-known components of medication packaging systems. The effects they have on product and patient safety partly depend on which patients, when, and through what delivery system. Aerosols, sprays, and transdermal packaging are high risk compared to tablets, capsules, or topical powders. Andrew Feilden, European E&L strategic director, Element Materials Technology, when looking back on how some regulations were formed, said, “Inhalation aerosols and sprays are deemed to have the highest likelihood of packaging component-dosage form interaction and the highest degree of concern based on the route of administration. As such, it is not too surprising that most of the early work on regulatory requirements focused on orally inhaled and nasal drug products. It was groups such as the International Pharmaceutical Aerosol Consortium on Regulation & Science that initiated and built on supply chain interaction and understanding” (2).

There is an interplay between the permitted concentration limit (PCL), the maximum daily dosage (MDD), and product material design. It is interesting to consider predictive modeling versus exposure for E&L and how far the industry has come with artificial intelligence (AI) in advancing predictive modeling and where the limits lie.

Over the past few years, there has been an increased prevalence for products to be classified as drug-device combination products. This classification leads to several additional considerations and challenges, not least with E&L study design and regulatory review of these studies. According to Nicholas Morley, principal scientist at Element Materials Technology, “Regional differences can exacerbate this and make it even more challenging for products seeking registration in multiple markets. Extractable and leachable studies performed on medicinal products typically follow standards and guidelines set out by the Product Quality Research Institute (PQRI) and the US Pharmacopeia (USP), whereas studies performed on medical devices follow standards set out by the International Organization for Standardization (ISO). Depending on the device type, there may be requirements to meet multiple different standards and guidelines which have similar aims (i.e., determining patient risk due to the product) but use different approaches to achieve them.”

No pun intended, this compounds an already complex process when trying to set minimums and maximums for E&L testing, particularly for high-potency drugs. Morley continues, saying, “When designing and performing studies on drug-device combination products, it is crucially important to have a deep understanding of the similarities and differences between the various extractable and leachable standards and guidelines, as well as the different regional expectations, and review authorities when designing E&L studies to minimize potential deficiencies and delays in approval.”

Where possible, the industry is accelerating migration toward continuous manufacturing and single-use systems (SUS). With regard to E&L, setting baselines and comparing differences can be helped by standardized protocols, which can facilitate “like-for-like assessments between each supplier’s SUS products and components” for end-users of SUS, according to Dan Rosen, vice-president and general manager of bioproduction at Thermo Fisher Scientific (3).

As said earlier, leachables being a subset of extractables, extractables testing can show which leachables might emanate from the material under standard conditions, or extreme conditions, and through simulation or modeling. However, one wrinkle to pay attention to is that new or different leachables might form during sterilization, storage, or use (3).

There is talk about a migration away from the well-characterized and established response of materials to gamma irradiation versus (due to lack of gamma testing capacity) employing less understood X-ray sterilization methods. “Current theories suggest that the E&L profile should be similar after X-ray or gamma sterilization, but it is important to perform extractables testing, says the Bio-Process Systems Alliance (BPSA)” (3).

According to Lalit Saxena, senior director of MSAT at Samsung Biologics, “Regulatory requirements are becoming increasingly stringent with respect to patient safety, and thus more in-depth risk assessment to implement single-use technologies for late-stage downstream unit operations is required” (4). Saxena pointed to the European Union’s Annex 1 as an example of regulatory expectations. Challenges are also being created by compliance with 

 <85> (for endotoxin certification) for single-use materials. Customized single-use assemblies increase the complexity, especially with components from multiple vendors, according to Saxena (4).

The impact on E&L study design can be easily overlooked. To understand this, it is important to determine who is responsible for the review of the extractable and leachable studies.

“Different E&L guidelines apply to pharmaceutical container closure systems (e.g., USP or PQRI) compared to medical devices (ISO). In additional, the different regional organizations can have their own specific requirements or interpretations. Extractables studies on container closure systems and single-use manufacturing process components often employ binary solvent systems,” states Morley. “Whereas CDRH [the Center for Devices and Radiological Health] prefers extractable studies to be performed using pure solvents. When a product is classified as a drug-device combination product (e.g., a pre-filled syringe containing an active drug-product), extractable studies may need to meet the requirements of the pharmaceutical container closure guidelines as well as the medical device standards simultaneously. Within the EU, if a drug-device product is classified as integral as is the case for pre-filled syringes, a notified body opinion is required prior to receiving a MAA [marketing authorization applications] approval. When designing and performing studies on drug-device combination products, it is crucially important to have a deep understanding of the similarities and differences between the various extractable and leachable standards and guidelines,” he concludes.

One of the great new tools for pharmaceuticals is the rise and integration of AI models and programing. Opinions vary in degree of enthusiasm, with many highly supporting greater inclusion, implementation, and reliance on such systems. But there are minority reports circulating among the E&L expert community. When asked how far the industry has come with AI in advancing predictive modeling for E&L, Rick Reiley, technical director, Extractus, replies, “We have not come far at all.” He points out that, “this is about the data sets available to train AI. The first problem is the data itself: data needs to be marked up with mass assignments, and modern software suites (Masshunter, UniFi/masslynx, and xcalibur) can do this. [However,] it is important to have correct identification of compounds as chromograph systems can vary the elution profile (ideally, compound assignments should be confirmed with a standard or reference material which are not always commercially available).”

Reiley continues to draw attention to a lack of extractable data sets that directly link to leachable data sets.“The vast majority of extractable data sets,” emphasizes Reiley, “do not have a corresponding leachable data set to train the AI, and those that do exist are not available for AI teams as these are company secrets. If you make a product and you disclose data to a regulatory body, the data [are] flat (no data files, just summary data and chromatograms/spectra) as giving more information invites greater scrutiny. If an AI algorithm were to find additional peaks from raw data, it would trigger review of a product license and potentially pull a product from the market.”

While on this subject, Reiley goes on to discuss a third hurdle regarding extractable data sets. “Very few have good data (meaning that peaks are not identified confidently, performed with rigor [usually single extraction with single injection] or separated chromatographically). There are problems with sensitivity limit of quantification (LOQ) that is not low enough), poor chromatography, poor identification (either none or letting the library decide) as well as inappropriate separation technique so the peaks are lost in noise or solvent peaks,” he states.

This last hurdle could be overcome by original equipment manufacturers (both instrument and packaging/material vendors) “as they don’t directly market drugs/medical devices, but they have incentives to avoid the research. You will have to source the materials for an extractable and leachable study, but it would need to be a large data set. Ideally, you would want to pick something with multiple suppliers (you don’t want to alienate a potential or current client), and it would require a lot of guesswork as E&L studies start with a risk assessment (calculating analytical evaluation threshold [AET] will vary depending on a number of factors). The alternative would be an independent body or an academic research group, but it would require a lot of funding (industry would not fund this) as E&L studies are expensive due to instrumentation and time required,” concludes Reilly.

Traditional analytics for identifying extractables and leachables are liquid or gas chromatography combined with mass spectroscopy (LC–MS or GC–MS). But according to Fran Adar, principal Raman applications scientist, Horiba Scientific, “From my discussions with Mark Witkowski [at FDA], I understand that the standard LC–MS and GC–MS techniques have difficulty identifying inorganic materials. In contrast, Raman spectroscopy is equally active for inorganic and organic compounds” (5).

Adar went on to say, “What Raman can do that those standard techniques cannot is identify minute amounts of material—typically 1–2 μm2 × 1–5 μm deep. And there are no waste products from the analysis that need to be disposed of. Is the information the same? Maybe yes and maybe no. The measurement is done on intact material whereas MS tears the molecule apart. There may be cases where information is lost in tearing the molecule apart” (5).

It could be that new tools from other application areas may migrate over to help out the E&L field before too long. There is certainly interest there, and some room for improvement.

Panzade, P. Nitrosamines, Elemental Impurities, Extractables and Leachables: A New Normal for the Pharma Industry. Webinar. Catalent. 

Feilden, A. Material Management and the Impact on Extractables and Leachables,” 

Markarian, J. Analyzing Extractables and Leachables in Single-Use Systems. 

Challener, C. Benefiting from Single-Use Tech Downstream. 

Adar, F. Exploration of the Use of Raman Microscopy to the Identification of Extractables and Leachables from Polymeric Containers. Spectroscopy 2022 37 (11) 8-13. DOI: 

Chris Spivey was previously the editorial director of 

When referring to this article, please cite it as Spivey, C. Testing the Limits of Extractables and Leachables. 

What are the lessons gleaned from a product’s E&L lifetime?