Feliza Mirasol is the science editor for 

Feliza Mirasol

Unidentified microbiologist is place the petri dish and tubes into incubator to incubate the culture of bacteria or fungi growth in appropriate condition, concept of microbial laboratory. | Image Credit: © Sukjai Photo – © Sukjai Photo – Stock.Adobe.com

Maintaining a sterile cleanroom environment and stringent cleanroom protocols in biopharmaceutical manufacturing are critical to ensure optimal product quality and, ultimately, patient safety. With the shift toward personalized medicines, new generation therapies, such as messenger RNA (mRNA), cell and gene therapies (CGTs), antibody-drug conjugates, and modern vaccines, are presenting sterility challenges. Therefore, it is becoming increasingly necessary for analytical technologies to advance and keep pace with the evolving, modern-day therapeutic requirements for sterile environments in cleanrooms.

As therapeutic modalities evolve, so do the challenges and methodologies associated with maintaining and verifying cleanroom sterility. Cleanroom standard operating procedures (SOPs) are the foundation to ensuring controlled, contaminant-free environments. Recent regulatory updates have prompted revisions to these procedures to align with contemporary standards and technological advancements.

For example, the International Organization for Standardization (ISO) updated the ISO 14644 series in 2015 (1), which delineates cleanroom classifications and associated protocols. In the series, ISO 14644-1 focuses on classifying air cleanliness by particle concentration, and ISO 14644-2 enumerates the requirements for testing and monitoring to ensure ongoing compliance with designated cleanliness levels. With these updates, a more rigorous approach to contamination control is emphasized, which necessitates that SOPs incorporate enhanced monitoring and maintenance strategies (1).

FDA, meanwhile, has advocated for integrating rapid and advanced detection methods into sterility testing, with the goal of expediting detection of microbial contaminants and reducing product release times without compromising safety. As a result, SOPs now often include provisions for implementing rapid microbiological methods (RMMs) alongside traditional testing techniques (2).

Sustaining sterility within cleanrooms poses multifaceted challenges, both enduring and emerging. Challenges that continue to persist include:

 Personnel remain the primary source of contamination. This is commonly attributed to inadequate gowning procedures, lapses in protocol adherence, and insufficient training—all of which can introduce particulates and microbes into the cleanroom environment (3).

Continuous surveillance of particulate and microbial levels is crucial; however, traditional monitoring methods may not provide real-time data, which potentially allows contaminants to proliferate before detection (4).

 Suboptimal cleanroom design or maintenance can lead to contamination risks. Improper airflow, inadequate filtration systems, and poorly maintained equipment are factors that can compromise sterility (5).

Meanwhile, emerging challenges posed by new therapeutic modalities, such as CGTs and mRNA-based treatments include:

 These new modalities often involve living cells or delicate nucleic acids, which are highly susceptible to environmental conditions. Therefore, maintaining appropriate temperature, humidity, and aseptic conditions is paramount.

 Production of these modalities typically involves intricate procedures, which increases the potential for contamination at multiple stages of the manufacturing process. Each step necessitates stringent controls to preserve product integrity.

 Many advanced therapies have limited shelf lives, necessitating expedited manufacturing and testing processes. Traditional sterility testing methods, which can take up to 14 days, are impractical for these products, prompting the need for rapid testing alternatives (6).

On the other end of contamination prevention is verification of sterility, which is also a critical step to ensuring a product’s safety and efficacy. Verification requires analytical testing methods, including membrane filtration, direct inoculation, and automated culture systems.

In membrane filtration, product is passed through a filter that captures potential contaminants. The filter is then incubated in growth media to detect microbial presence (7). Direct inoculation is applied to products that cannot undergo filtration. In this method, product is directly inoculated into culture media, incubated, and microbial growth is monitored over a specified period of time (8). Meanwhile, automated culture systems were developed to enhance efficiency and reliability. In these systems, product is continuously monitored for microbial growth. This method offers advantages in sensitivity and reduced hands-on time compared to manual methods (9).

The evolving regulatory requirements over the years as well as the increasing biopharmaceutical complexity and growing demand for aseptic processing have prompted significant advancements in cleanroom technologies and analytical testing methods. These innovations have been driven by the need for faster, more accurate contamination detection, improved environmental monitoring, and greater flexibility in biomanufacturing. Innovations include rapid microbiological methods (RMMs), next-generation sequencing (NGS), real-time environmental monitoring, and the use of digitalization and artificial intelligence (AI) in cleanroom analytics.

The adoption of RMMs has been prompted by the lengthy time it takes for traditional sterility testing methods. For example, membrane filtration and direct inoculation can take up to 14 days to get results. RMMs utilize technologies such as nucleic acid amplification (e.g., polymerase chain reaction-based assays), flow cytometry, and adenosine triphosphate bioluminescence to detect microbial contaminants in real-time or within hours rather than days (4). Advanced therapies such as CGTs and mRNA-based therapeutics require faster contamination detection than traditional methods and have been the real drivers behind accelerating the implementation of RMMs.

NGS, meanwhile, has emerged as an important tool for microbial identification in cleanrooms as it enables the detection and characterization of diverse microbial populations, unlike conventional culture-based methods. NGS also enables the detection of previously unculturable organisms. This technology enhances contamination tracing and root-cause investigations and can lead to more effective contamination control strategies (10).

Real-time environmental monitoring has been enabled with the development of advanced sensors. The ability to monitor particles and microbes in real time is changing the way contamination control is handled in the cleanroom. For example, technologies such as laser-induced fluorescence sensors allow for immediate airborne microbial detection without the need for culture-based methods. Systems such as those are often integrated into continuous environmental monitoring platforms and provide instant alerts to deviations in cleanroom conditions (2).

Finally, AI-driven analytics, combined with machine learning algorithms, are being deployed to analyze environmental monitoring data, detect contamination patterns, and predict contamination risks before they occur. Having this predictive approach enables proactive decision-making and reduces the risk of contamination events, which in turn minimizes production shutdowns (11).

Cleanrooms and Associated Controlled Environments Standards

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2. Thermo Fisher Scientific. Rapid Sterility Testing: Overcoming Challenges in Cell Therapy Production. 

Cleanroom Technology: Fundamentals of Design, Testing and Operation

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4. Agalloco, J.; Akers, J. 

Advanced Aseptic Processing and Biopharmaceutical Manufacturing

Cleanroom Microbiology for the Non-Microbiologist

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6. Jallow, J. Rapid Sterility Testing for Cell and Gene Therapy Treatments. 

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9. Khuu, H. M; Stock, F.; McGann, M.; et al. Comparison of Automated Culture Systems With a CFR/USP-Compliant Method for Sterility Testing of Cell-Therapy Products. 

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10. Xiao, Y.; Zhang, L.; Yang, B.; et al. Application of Next Generation Sequencing Technology on Contamination Monitoring in Microbiology Laboratory. 

 (1), 25–31. DOI: 10.1016/j.bsheal.2019.02.003
11. Bradshaw, N. Using AI and Machine Learning in Cleanroom Contamination Control. 

®
Vol. 49, No. 4
May 2025
Pages: 28–29

When referring to this article, please cite it as Mirasol, F. Evolving Cleanroom Procedures and Sterility Challenges. 

Articles

The shift toward personalized medicines poses new challenges in cleanroom protocols.

Evolving Cleanroom Procedures and Sterility Challenges

On April 24, 2025, CN Bio, a provider of organ-on-a-chip (OOC) systems and solutions aimed at accelerating drug discovery and development workflows, announced that it has established a strategic partnership with Pharmaron, a China-based R&D service provider for the life sciences industry, under which Pharmaron will validate CN Bio’s PhysioMimix technology across existing applications and will integrate OOC technologies into its R&D platform. Also under the deal, the partners will explore the development of new applications that address unmet needs in drug discovery and development.

In the initial phase of the collaboration, the companies will focus on validating CN Bio’s PhysioMimix technology in current applications of disease modeling, toxicity testing, and absorption, distribution, metabolism, and excretion (ADME) studies. Upon successful validation, the companies will then aim to adopt the platform in priority R&D areas while co-developing novel applications to expand the capabilities of OOC technologies.

“Pharmaron [is] the ideal partner for CN Bio as we look to accelerate the growth of our OOC solutions portfolio in new and existing application areas. By working closely together and leveraging the [expertise] of each company, we can share resources that enable us to progress new, innovative solutions from concept to global market deployment more effectively than ever before,” said Paul Brooks, PhD, CEO of CN Bio, in a company press release (1).

As a global drug R&D service platform, Pharmaron specializes in providing end-to-end services across drug discovery, preclinical, and clinical development. Through this partnership with CN Bio, Pharmaron will install PhysioMimix instruments at its global facilities to enable the joint development of cutting-edge OOC solutions to meet evolving R&D challenges.

“The developments this partnership will enable are especially important given the recent FDA announcement, outlining their plan to phase out animal testing requirement for monoclonal antibodies and other drugs with more human-relevant models. Our platform is best placed to serve current market gaps including testing for immune-mediated organ damage. Together we can expedite this key development in line with global momentum for more ethical advancements that reduce costs and improve human health,” said Tomasz Kostrzewski, PhD, chief scientific officer, CN Bio, in the release. FDA made its 

 about phasing out animal testing requirements earlier in April (2).

Over the past several years, manufacturers have been developing OOC technology, largely consisting of engineered or natural tissues grown inside microfluidic chips, which allows them to maintain tissue-specific functions, according to a 

 (3). According to the article, OOCs today encompass an array of tissue and organ functionalities, including the blood-brain-barrier, central nervous system, heart, intestine, liver, lung, kidney, muscle, and skin, which allows pharmaceutical companies to better understand the underlying pathophysiology of diseases as well as determine the effect of drugs 

 (4). Market research predicts that the global OOC market will be worth $388 million by 2028, up from $82 million in 2023, exhibiting a compound average growth of 36.4% from 2023 to 2028 (5).

“We are committed to continuous innovation that enhances the quality of our services and accelerates drug discovery and development. Organ-on-a-chip technology holds significant potential to advance translational science by improving our understanding of drugability at the preclinical stage. We are pleased to collaborate with CN Bio to explore how this technology can add meaningful value to our partners’ development programs,” said Hua Yang, PhD, chief scientific officer, Pharmaron, in the company press release (1).

CN Bio and Pharmaron Establish Long-Term Strategic Partnership to Develop OOC Technologies on a Global R&D Platform

. Press Release. April 24, 2025.
2. FDA. 

FDA Announces Plan to Phase Out Animal Testing Requirement for Monoclonal Antibodies and Other Drugs

. Press Release. April 10, 2025.
3. Barton, C. 

 (5), 16–17, 30.
4. Leung, C. M.; de Haan, P.; Ronaldson-Bouchard, K.; et al. A Guide to the Organ-on-a-Chip. 

Global Organ-on-Chip Market Size, Share, Trends, COVID-19 Impact and Growth Analysis Report—Segmented by Type (Heart-on-chip, Human-on-chip, Intestine-on-chip, Kidney-on-chip, Liver-on-chip and Lung-on-chip), Application and Region (North America, Europe, Asia-Pacific, Latin America, Middle East and Africa)—Industry Forecast from 2023 to 2028

Under the partnership, Pharmaron will validate and promote the application of CN Bio’s PhysioMimix technology and will integrate OOC technologies into its R&D platform.

CN Bio Forms Strategic Partnership with Pharmaron to Develop OOC Technologies 

This episode of Drug Digest will discuss where the significant bottlenecks are in the development and manufacturing of new molecules resulting from progressive technologies, such as AI and ML, and how best to approach these challenges.

Keeping Pace with Progressive Technologies in Biopharma Development

The biopharma industry is at a time where progressive technologies such as artificial intelligence, machine learning, and digitalization technologies are making a significant impact on how drug discovery, development, and manufacturing are being driven. Yet the question remains as to how the industry itself is keeping up with the fast-paced innovation churning out from these advanced technologies. This episode of 

 will examine where the significant bottlenecks are in the development and manufacturing of new molecules resulting from these progressive technologies, and how best to approach these challenges.

Kevin Dondarski, Principal, Life Sciences R&D, Deloitte Consulting

Maryland Franklin, PhD, Vice-President and Enterprise Head, Cell and Gene Therapy, Labcorp

Lidia Serina, PhD, head of Development Services at Simtra BioPharma Solutions

Sean Tucker, PhD, Founder and Chief Scientific Officer, Vaxart

® editors, who interview industry experts to discuss the emerging opportunities, obstacles, and advances in the pharmaceutical and biopharmaceutical industry for the research, development, formulation, analysis, upstream and downstream processing, manufacturing, supply chain, and packaging of drug products.

Developments in Small-Molecule APIs, Excipients, and Ingredients

Videos

Drug Digest: Keeping Pace with Progressive Technologies in Biopharma Development

Discussion about advancements in flow sensor technologies with PSG Biotech's Michael Franco.

INTERPHEX 2025: Advanced Flow Sensor Technologies Enable More Accurate Measurement

At INTERPHEX 2025, Michael Franco, global sales director at PSG Biotech, spoke with the PharmTech Group about how recent advancements in pharmaceutical flow sensor technology focus on improving measurement accuracy as drug production becomes more expensive and precise. A key development, he notes, is the enhancement of Coriolis single-use flow meters, which now feature a higher turndown ratio. This enhanced feature allows a single sensor to accurately measure both high and low flow rates. With this innovation, inventory needs can be reduced, and sensor usability across various process conditions is extended, which improves sustainability.

“When we talk about high turndown ratio, it's very good for the industry, because with one sensor, you can measure more than just low flows or high flows. So, in terms of sustainability, there's less need for inventory, and the sensor can be used for longer, because it can be operated in different process conditions. [In] terms of the future, we're adding more digital communication platforms so customers can communicate to the sensors in different ways, making sure that the sensors talk to pumps, and to get a really good performance between the two. [Those] are the things that probably are going to be asked more and more from customers,” Franco says.

Looking forward, this integration of digital communication platforms to enhance sensor interoperability is expected to enable better coordination with pumps and optimize system performance. These advancements are increasingly demanded by customers for improved bioprocessing applications, including tangential flow filtration (TFF) and chromatography skids. The ongoing adoption of these technologies as observed at INTERPHEX, notes Franco, highlights their growing importance in pharmaceutical manufacturing.

INTERPHEX 2025 occurred in New York City on April 1–3.

Michael Franco, Global Sales Director, PSG Biotech

Michael Franco leads the PSG Biotech global sales team, focused on strategic business growth, with a strong focus on adding value to PSG Biotech customers, and building meaningful and in-depth partnerships. Mike’s abilities to connect with customers and understand business strategy stem from a rich educational and career experience in key roles in multiple industries:

Prior to Biopharma, Mike worked in several sectors including Aerospace, Industrial, Renewable Energy and Power. He progressed from positions such as Researcher, Applications Engineer, Commercial Engineer, Segment Leader and Regional Sales Manager where he excelled and elevated to his current position as Director of Global Biopharma Sales. Companies that have been graced with Michael’s employed expertise include Syntest Corporation, Panametrics, GE Power Systems, GE Oil and Gas, GE Industrial, GE Energy Services, and AMETEK.

Michael’s education achievements include a Bachelor of Applied Science in Electrical Engineering from Boston University, a Master of Science in Information Technology at Worcester Polytechnic Institute, and a Master of Business Administration with a concentration in Business Administration and Management at the Isenberg School of Management at UMass Amherst.

Michael Franco, global sales director at PSG Biotech, spoke with the PharmTech Group at INTERPHEX 2025 about recent advancements in pharmaceutical flow sensor technology.

Discussion with WMFTS' Nicole Hunter about the industry initiative around single-use assembly components caused by supply disruptions from the COVID-19 pandemic.

INTERPHEX 2025: Industry Initiates Interchangeable Single-Use Assembly Components in Response to COVID-19 Supply-Chain Disruption

During the COVID-19 pandemic, supply-chain disruptions led to extended lead times, sometimes exceeding a year, which significantly impacts biopharmaceutical manufacturing and patient access to therapies, says Nicole Hunter, head of Global WMArchitect at Watson-Marlow Fluid Technology Solutions (WMFTS). Hunter spoke with the PharmTech Group at INTERPHEX 2025, which occurred April 1–3 in New York City.

“We really saw in COVID this really exacerbated example of demand exceeding supply. What we saw was lead times going up for assemblies to sometimes [more than] 52 weeks, which is [a] really, really long time and obviously very disruptive for the biopharma company and getting the therapy to patient[s]. So, one of the reasons for that was that one component would be validated in an assembly, and that meant if that component was difficult to get hold of—because the demand was really, really high—you couldn't supply the whole assembly,” Hunter says. “So, what would be better is if you could have that component or another one built in, where the other component may be more freely available, and then you can minimize the time to deliver the whole assembly and minimize the impact to patients.”

As Hunter notes, the dependence on specific validated components in assemblies was a key issue. This dependence caused bottlenecks when those components became scarce due to high demand. To address this challenge, the bio/pharmaceutical industry began an initiative and introduced the concept of interchangeable parts. This approach allows for substituting validated components with equivalent alternatives that meet the same specifications in terms of fit, form, and function, Hunter explains.

By pre-agreeing on the equivalency between suppliers and manufacturers, the process remains compliant, maintains quality, and minimizes disruptions to production workflows. This initiative enhances supply chain resilience by reducing dependency on single-source components and improving assembly efficiency, Hunter emphasizes. She points out that industry professionals are discussing advancements in this field and looks forward to the collaborations and innovations that will be fostered at INTERPHEX to optimize bio/pharmaceutical manufacturing processes.

Hunter gave a talk at INTERPHEX 2025 on “

Futureproofing Single-use in Bioprocessing

Nicole Hunter, PhD, Head, Global WMArchitect, Watson-Marlow Fluid Technology Solutions

Nicole Hunter, PhD, is the head of WMArchitect single-use solutions at Watson-Marlow Fluid Technology Solutions (WMFTS). Prior to joining WMFTS, she held the position of R&D manager at Thermo Fisher Scientific leading a team of chemists and microbiologists developing 

 diagnostic devices. Her PhD in the area of pharmaceutical materials science was undertaken at the University of East Anglia, United Kingdom. Hunter also has pharmaceutical industry experience gained from working in the drug discovery department of Pfizer Animal Health. She is an active member of several industry working groups and committees including BioPhorum, the Bio-Process Systems Alliance, the International Organization for Standardization, and the British Standards Institution.

Sitting down with the PharmTech Group at INTERPHEX 2025, Nicole Hunter, head of Global WMArchitect at WMFTS, explains the industry initiative around single-use assembly components resulting from supply disruptions caused by the COVID-19 pandemic. 



ITT's Dave Loula speaks about innovation in advanced sensing technologies that answer customers' need for reliable valve function at INTERPHEX 2025.

INTERPHEX 2025: Innovations in Advance Sensing Technologies that Answer the Need for Reliable Valve Function

The primary focus in valve technology advancements is reliability, particularly in pharmaceutical applications where system integrity and contamination prevention are critical. Talking with the PharmTech Group at INTERPHEX 2025, Dave Loula, global product director, ITT Engineered Valves, emphasizes that customers seek valves that are easy to maintain and provide consistent performance. Key innovations in advanced sensing technologies are those that address customers’ need for reliable and easily maintained valves.

“The advanced sensing—we term this ‘Integrated Sensing Platform,’ our new ISP technology—is part of that reliability solution that the customers are looking for when they're looking for feedback on their valves. They want to make sure that they're getting a reliable sensing of the position of the valve so that they know that a valve is in the open position or in the closed position so they can go on and do their particular operation. The ISP, which is a very small, compact device that fits on top of one of our valves, provides that type of reliable feedback on valve operation,” Loula says.

ISP technology plays a crucial role in maintaining process integrity, reducing the risk of system failures, and ensuring compliance with stringent regulatory requirements. Additionally, the broader platform includes sensors, actuation systems, and advanced monitoring solutions to support end users, original equipment manufacturers, and distributors, Loula explains.

Loula is most looking forward to engaging with industry stakeholders at INTERPHEX to foster collaboration and knowledge exchange. He believes that such engagement ensures that emerging needs in pharmaceutical valve technology are met with cutting-edge solutions.

Attendees can visit ITT at Booth #3337 at INTERPHEX 2025, being held in New York City on April 1–3.

David Loula, Global Product Director, ITT Engineered Valves

David Loula has extensive experience in the biopharmaceuticaland industrial sectors, specializing in sales,product management, and business strategy. Currently serving as global product director and previously as global marketing manager at ITT Engineered Valves since August 2012, Loula has driven significant product initiatives and marketing strategies. Prior roles include business development manager at ITT Industries and sales manager at ITT PureFlo, where he managed sales efforts for the Pure-Flo hygienic diaphragm valve product line across a 15-state region. Loula's educational background includes a Bachelor of Science in Mechanical Engineeringdegree from the University of Michigan.

Speaking with the PharmTech Group at INTERPHEX 2025, Dave Loula, global product director, ITT Engineered Valves, discusses innovation in advanced sensing technologies that addresses customer need for reliable valve function.