What are some general industry trends/issues that pharm/biopharma companies are dealing with in regards to microbiology testing, analysis and identification?
Alan Hoffmeister, Global Field Technical Manager, Charles River, Microbial Solutions and Jeremy Robertson, Celsis® Senior Product Manager, Charles River, Microbial Solutions: A major trend among pharmaceutical companies today is the movement from traditional microbiology detection methods to rapid micro detection methods. Traditional microbiology methods require days to weeks to confirm the absence of microbial contamination. With current pressure from the industry to streamline manufacturing processes, pharmaceutical manufacturers are looking to rapid micro methods to quickly release product from micro hold. The implementation of a rapid micro method enables quality assurance in less than half the time of traditional methods. The result is reduced production time, lower costs to manufacture, reduction in inventory and safety stock, and quick response to contamination events. Not only do these results lead to significant financial savings, but they also allow faster response to market demands.
Lakiya Wimbish, Product Manager for Endotoxin Testing, Lonza: One key issue in the field of bacterial endotoxin testing is how to deal with Low Endotoxin Recovery (LER) where the presence of endotoxin is ‘masked’. Two common drug excipients, polysorbate and citrate, have been identified as probable causes. However, the industry is divided over how to deal with the issue of LER.
While purified lipopolysaccharide (LPS) preparations such as Control Standard Endotoxin (CSE) may be masked under certain conditions, the LPS in Naturally Occurring Endotoxin (NOE) may be protected from the dispersing effect of chelating buffers. Some in the endotoxin testing community believe NOE should be adopted for hold-time studies. However, others favor demasking protocols to detect LPS in LER conditions. There is also an unanswered question regarding the risk to patient safety through immunological effects of pyrogens.
A second key issue is the introduction of alternative assays, such as the PyroGene™ recombinant Factor C (rFC) Assay and Monocyte Activation Test. These tests are intended to reduce the industry’s reliance on the horseshoe crab. Although regulatory bodies have expressed formal support for use of alternative methods, a simple, extra validation process is required for implementation.
Cheryl Zaman-Zadeh, Director of Sales at Novatek International: With regulatory trends moving to requiring risk-based quality programs, the sampling for microbial contaminants must reflect the risk assessment performed on the process. Sampling programs will move from the traditional grid based approach to a risk-based approached.
Analyzing the data generated from the test results will also receive added regulatory scrutiny. The integrity of the data will be looked at closely making manual programs, prone to human error, obsolete for monitoring manufacturing processes. Software based programs will be required to be compliant.
Dr. Gilberto Dalmaso, Global Aseptic Processes Development Manager, Life Sciences Division, Particle Measuring Systems:
- Quality Risk Management
- Microbiological Data Integrity
- Microbiological Methods Validation
- Environmental Monitoring
Can you tell us about some new technologies and/ or processes that are having a noticeable impact on pharma/biopharma company’s microbiology testing and analyzing operations?
Hoffmeister and Robertson: Rapid microbiological methods (RMM), such as cartridge-based technology for quantifying endotoxin and the amplified ATP bacterial detection systems, have contributed to the advances being seen in the microbiology labs of the pharma/ biopharma industries. The rapid endotoxin detection cartridge system allows for in-process control testing to be performed at the point of sample, within 15 minutes, removing the need for at-risk process continuation and allowing the industry to better adopt Quality by Design (QbD) principles. The rapid amplified ATP bioluminescence detection system has reduced the potential hold time for finished product from weeks down to days, allowing product to reach the market much sooner.
Rapid bacterial identification via MALDI-TOF mass spectrometry has also entered the pharma/biopharma industries, and is providing faster, more accurate bacterial identification for product testing and environmental monitoring as compared to traditional phenotypic methods and sequencing. This enables quicker investigations to be carried out and faster decisions to be made about product and environments, which can be vital to ensuring patient safety.
Patrick Hutchins, PhD, Global Product Manager | Biotechnology, TSI, Inc: For the entire history of aseptic manufacturing, the only practical metric of airborne microbial contamination has been the colony forming unit (cfu). Adapting growth-based approaches for monitoring modern aseptic manufacturing processes is proving difficult. Although microbiologists have bemoaned the limitations of measuring only cfu, no alternative metric or technique has been embraced. One alternative microbiological method, called Laser Induced Fluorescence (LIF), is showing potential to actually replace compendial methods for inprocess environmental monitoring applications. LIF is not growthbased. Rather, it detects individual microorganisms in real-time, giving it the flexibility needed to improve aseptic manufacturing processes. By eliminating interventions for agar plate changes and by providing real-time, continuous data, LIF can reduce risk while increasing manufacturing efficiency and capacity in certain settings.
Wimbish: In endotoxin testing, data analysis is a considerable challenge, especially in terms of getting data into the core information system. Currently, a significant number of laboratories manually transcribe their results into their information management system. Not only is this laborious, but it is also a very error-prone process. Software solutions such as Lonza’s WinKQCL™ and MODA™ Software provide built-in features that automatically transfer data from one system to another - offering data analysis time savings. By showing a simple pass/fail flag for critical pieces of data, data review is simplified, while a comprehensive trending analysis feature offers the ability to quickly generate larger reports for more complex analysis.
Increased adoption of automation in QC testing is helping pharmaceutical companies to improve efficiency, reduce human error and consequently make cost savings. Robotic systems can be used to automate the preparation of the samples and controls and add working reagent to the plate. Software solutions further improve efficiencies when interfaced with robotic systems and can also allow companies to move away from paper-intensive QC analysis. To help laboratories better understand the benefits of automating QC testing, Lonza has been running justification workshops at customer sites to analyze the potential return on investment.
Zadeh: New applications of computers have made environmental monitoring programs including microbial testing more efficient and cost effective than ever. The use of Cleanroom-ready tablet computers, scanners, and instrument connectivity now allow for real-time data capture in classified areas while securing the sampling process. This combined with environmental monitoring software allows for complete traceability from acquisition to identification of micro-organisms. This combination of the latest software and hardware has revolutionized the way monitoring is executed. These technologies will further provide compliance to the new FDA regulations on Data Integrity.
Dalmaso: Real Time Environmental Monitoring and Rapid Surface Microbiological Monitoring
- Environmental Monitoring Data Management (including microbiological data)
- VHP decontamination processes
In the past few years have regulatory expectations from the FDA and other global agencies resulted in more scrutiny placed on microbiology processes? Can you provide any specific details?
Wimbish: For the production of biologics, the occurrence of LER in hold-time studies has concerned regulatory authorities worldwide. Although there is no data to support that this poses a patient safety issue, the FDA is particularly cautious. This has driven manufacturers to rethink how they formulate their biologics and vaccines to avoid LER-prone buffers.
Since the introduction of the FDA’s Quality by Design (QbD) approach, greater emphasis has been placed on pharmaceutical manufacturing processes. QbD is based on the principle that quality can’t be tested into products, it should be built-in or by design. Implementation of QbD drives improvements in production cycle time, efficiency, cost and product quality.
QbD encourages more raw material and in-process testing to better understand the manufacturing process, highlighting where adulterations are most likely to occur. It also provides a means of identifying contamination early on before the final product stage, allowing for both time and cost savings.
Recently, the FDA has updated guidelines in USP Chapter 161 to provide manufacturers with a standardized set of regulations for the proper implementation, measuring and documenting of tests for monitoring endotoxin levels on their products. While these updates are relatively minor, it is important that they are adhered to.
Hoffmeister and Robertson: The advent of the highly accurate, rapid microbiology system has meant that the scrutiny levels placed on traditional microbiological methods has increased. Subsequently, regulators are expecting to see a higher level of proof as to the accuracy of results being reported than traditional methods can provide. Another example being pressed is trending data of the identified isolates recovered from production facilities to demonstrate control of the environment to reduce the likelihood of contamination risk. Accuracy and reportable rates for identification of unknown isolates is becoming more important for biomapping facilities. Another example is the recurring discussions regarding visual inspections during media fills and sterility tests; more emphasis is put on challenging the ongoing qualification program for the operators for the error-prone evaluation of turbidity. Therefore, we observe a trend towards implementing more robust and objective technologies for this purpose, like amplified ATP bioluminescence.
Dalmaso: YES, see the list below of FDA observations in 2015. Most of them are related to microbiological processes. FDA-CDER-listing observations 2015 (microbiologicals are in bold):
References
What advice would you give to a pharmaceutical company struggling with their microbiology processes/applications? Is there a top 10 list of items you would suggest to these companies to help them?
Wimbish: For a pharmaceutical company struggling with their microbiology processes, we would recommend the following:
- Consider automation to improve efficiencies and to reduce human error.
- Commit to better training for employees. There are a wide range of tools out there. Ask your test suppliers to see what training they can provide, such as webinars and e-learning modules.
- Secure a secondary supplier or alternative methodology for products where LAL is not the optimal method for detection of endotoxins, including filters and plant-based products.
- Implement better data capture/analysis to ensure faster, more accurate and more complete data capture and rapid traceability should it be needed. Data analysis software that offers trending capability enables the end-user to review and analyze on demand, to quickly identify trends and respond before out-ofspecification errors occur. This is an important tool for regulated environments.
Dalmaso:
- Methods validation
- Air monitoring plan and methods (especially in ISO 5 environment)
- Risk Assessment
- VHP validation and use of Biological Indicators
- Surfaces monitoring plan and methods (especially in ISO 5 environment)
- Culture media reading and data integrity/management (ie. Environmental monitoring).
- Cleanroom operators certification program (especially in ISO 5 environment)
- Sterility Assurance programs
- New technology implementation strategy
- Regulatory lobby and meetings/relationship
What do you see as the major industry critical issues over the next five years in regards to microbiology?
Hoffmeister and Robertson: With biologics and biosimilars on the rise as an area of therapy, time is of the essence when it comes to releasing product. Due to shorter expiration dates as compared to traditional to small-molecule pharmaceuticals, some biologics may already be administered by the time traditional micro methods yield results. The ability to get a biological drug to market faster without sacrificing quality is paramount to ensure maximum treatment effectiveness. Furthermore, outbreaks such as the Zika and Ebola epidemics require pharmaceutical manufactures to scale up production rapidly to respond to the health crisis.
When it comes to cost, biologics are over 20 times more expensive than a small-molecule drug. With their increased demand, so comes increased pricing pressures for manufacturers. To stay competitive and keep pace, pharmaceutical companies will need to look for new ways to reduce their manufacturing costs. Using rapid micro methods is an effective and efficient way to shorten production cycle times, reduce manufacturing costs, expedite product time to market, and allow pharmaceutical companies to respond to immediate market demands.
Wimbish: The future of endotoxin testing will be influenced both by the sustainability of a natural resource and uptake of alternative endotoxin detection methods. The LAL assay is one of the few remaining assays reliant on an animal source (in this case, blood extracted from horseshoe crabs) for a key raw material. While recombinant endotoxin testing methods are available on the market, such as the PyroGene™ recombinant Factor C (rFC) Assay, their classification as ‘alternative’ tests by both European Pharmacopoeia (EP) and the Food and Drug Administration (FDA) have until now proven to be an obstacle to their large-scale adoption. Over the next five years, we expect the use of animal-free methods will be on the rise due to companies’ sustainability initiatives, particularly in the EU.
It is also likely we will see changes in the way we perform endotoxin testing in order to tackle the LER challenge. This may affect sample preparation techniques and we may even see the introduction of new test methods.
Dalmaso:
- Lack of innovation and scientifically sound methods of implementation
- Pharmaceutical microbiologist education/university programs (specific university plans especially in Europe, Asia and South America): In Europe/Asia/South America there aren’t any university pharmaceutical microbiology courses and we don’t have a pharmaceutical microbiologist in pharmaceutical companies playing key roles.
Hutchins: The evolution of aseptic manufacturing is trending towards increasingly isolated and automated processes. The growing adoption of isolators for fill finish and the emergence of continuous manufacturing processes are ready examples. Traditional methods of airborne microbial monitoring, i.e capturing and culturing microorganisms, are not adapting sufficiently. In certain settings microbial monitoring can introduce significant risk. Indeed, the manual manipulations associated with providing fresh growth media inside the aseptic core directly undermine efforts to isolate drug product from operators. Moreover, growth-based microbiological methods—no matter how rapid—cannot provide the real-time data necessary to support continuous or near-continuous processes. Over the next five years microbiological methods must adapt beyond inflexible growth-based approaches in order to support safer, more efficient manufacturing processes and technologies.