How to Audit Your Cryopreservation Workflow for Microbial Risks

 

Tubes of biological samples being placed in liquid nitrogen. Image by Dr. Vereczkey Attila CC BY-SA 3.0

Cryopreservation is an essential technology for pharmaceutical companies because it effectively preserves delicate biological samples. Storage systems are elaborate and sensitive, requiring specific temperature settings and no contaminants. Plenty of microbial risks could jeopardize everything from cells to organs, so knowing the best ways to audit workflows to eliminate these circumstances is essential.

By Emily Newton 

Review Current Floor Plans and Workflows

Workers must identify every potential contamination source by creating maps of stakeholders’ workflows. From fleet drivers to lab technicians, each step could introduce a new variable. All devices, personnel, and equipment must be reviewed at each stage to discover compliance and hygienic oversights.

 

During this stage, companies should also take stock of their most common microorganisms and discover ways to promote their survival. The facility may not be conducive, even in controlled cryovessels, to support the culture’s long-term health. This is why floor plan reviews are critical for finding bioreactor contamination situations, among other compromises. It is the reason many organizations fall short of compliance.

Mitigate Microbial Contamination Points

Observe the map and create a directory of potential contamination points. Everyone coming into contact with cryopreservation materials and the cells needs analyzing. Risks caused by human error, such as not wearing face masks or problems with the facility environment because of air quality, may also exist. Raw materials on tools and other machinery could also enter cryopreservation environments.

 

Pharmaceutical companies should perform these evaluations multiple times to understand the full scope of the workflow. For example, worker injuries may not happen every day, but a potential cut could expose specimens and transmit infection.

 

Workforces must challenge existing controls governing how to manage these samples and microbial influences and see how to improve. Staff must adequately sterilize all tools, using the most up-to-date disinfection protocols. Workers must maintain cryopreservation technologies to ensure their integrity. Personnel should also know how to document any activity regarding biological materials. If issues arise, then the source is more easily located.

Implement Microbial Testing and Observation

Most of these auditing steps concern processes before primary interaction with the microbes, ensuring temperatures range from -160 to -180 degrees Celsius. However, people working directly with biological materials must implement tests to avoid working with compromised in-process cultures.

 

Researchers should observe the quality of their clean rooms and environmental monitoring conditions, as well as the final cryopreserved product. This final observation phase could reveal unnecessary burdens or stressors on the microbes that others missed. Then, these professionals can validate the efficacy of previous quality assessments, validating if people are familiar and well-trained enough to discover something that could harm development.

Use Various Sterility Tests

Sterility testing is mandatory. Products must be free of microorganisms to preserve a drug’s effectiveness. Additionally, thorough decontamination and cleaning protocols avoid false positives when testing for contaminants, streamlining workflows further.

 

The direct inoculation method is reliable. It transfers samples into two tubes aseptically and lets them incubate at high temperatures. Membrane filtration works for cultures with properties that would not work in the conditions required for direct inoculation.

 

Experts should also ensure other protective measures are as effective as possible. Materials like dimethyl sulfoxide, which is a common cryoprotectant, are critical for preventing ice crystals in cells. Technicians should dry it out as much as possible for peak effectiveness by using one of several methods, including:

 

●     Distillation from CaH2

●     Molecular sieves

●     Vacuum distillation

Review Data and Find Knowledge Gaps

Experts need to assess the data from workflow testing for knowledge gaps. Perfect and flawed reports alike could contain insights. Many documentation processes reveal trends in how people handle the products, reinforcing or debunking a company's quality control and mitigation strategies. The company may believe it incorporates first-in, first-out methods, but it may not be thoroughly applied in practice.

 

What was once reliable could also become inefficient. Consistently performing these tests gives a comprehensive overview of historical performance, letting people know the first instances of failures in commonly compliant areas. Deviations are the first flags in making microbes healthier among stakeholders.

Use Corrective and Preventive Actions (CAPA)

CAPA addresses the gaps analysts discover in testing reports. For example, it could include using different materials to wash workstations or minimizing contact with transportation workers. Root-cause analysis will be a continuous theme at this stage of auditing. It unravels out-of-specification results and inspires process discovery to optimize operations.

 

It could also signal an opportunity to upgrade outdated equipment that is losing durability or consistency. Many cryogenic refrigerators and other containers incorporate smart technologies, which monitor temperature changes and notify operators of dangerous anomalies. They could also automatically divert to backup power resources during an outage or other emergency to ensure essential drugs are safe.

 

Frequently, CAPA holds organizations accountable and makes them more compliant with the most essential regulations, including the Federal Drug Administration, the International Council for Harmonization and the European Medicines Agency. Many mandate testing procedures and how to establish safe working environments. It could change how facilities filter water or distribute dosages between cryogenic containers.

Microbial Quality Control

Quality assurance in pharmaceutical research is crucial for ensuring customers stay healthy. Therefore, keeping cryopreservation resources contamination-free is one of these experts' most important jobs. If they succeed, new medicines could save lives and medical facilities will execute patient recovery faster.

 

However, auditing how workers interact with microbes in cryopreserved spaces requires patience and attentiveness. Organizations must implement these practices immediately to establish good reputations and ethical practices.

 

Pharmaceutical Microbiology Resources (http://www.pharmamicroresources.com/)

The Toxins Within Smoke and Soot

 Image: Soot by sunlight, by Tim Sandle.

We all have a basic notion of what smoke is. For example, maybe you have witnessed a home on fire. You see flames coming out of a window or a roof, but mostly, what you see are billowing clouds of black, grayish and even some white smoke.

 

After firefighters extinguish the blaze, the floors, furniture, ceiling, carpets and just about everything inside the structure will be coated in black soot.

 

The difference between smoke and soot is distinct. Simply put, smoke is that substance that is airborne and escapes upward into the atmosphere. Soot is solid matter that is deposited on physical objects.

 

Think of it this way: Smoke is what flies up a chimney and out into the air. Soot is that which sticks to the side of a chimney and can build up dangerously to pose hazards, such as a chimney fire. That’s why chimney sweeps are needed to clean away soot build-up on the interior surfaces of chimneys as a matter of routine maintenance.

 

So, smoke creates soot, although soot is also generated and deposited directly by a burning substance. Most people think of the byproduct of burning to be ash. However, there is a difference between ash and soot. Ash is defined as “inorganic, incombustible residue” that is generally gray or white in color but can also be black.

 

Soot, on the other hand, is most often black and sticky and manifests as a “coating” on surfaces. Ash is generally deposited on the ground or inside a burning device, such as a wood stove.

 

Toxic and Dangerous

 

Both soot and smoke are highly toxic substances.

 

For example, smoke from a house fire may contain up to 265 known cancer-causing agents. Smoke is made up of tiny particulate matter of wide variety. A combination of heat and atmospheric dynamics is what gives these billions of tiny particles to act as that flowing and amorphous “cloud” we identify as smoke.

 

Breathing in smoke is dangerous because the toxic particulates can be deposited inside the soft human tissues of lungs. That includes smoke from natural substances, such as trees in a forest.

 

However, smoke from man-made items tends to be far worse and more dangerous. Think of all the different chemicals and substances that make up, say, a chair or items of plastic. Both are comprised of dozens of different chemicals, synthetic substances along with natural fibers and other components.

 

Soot is also riddled with hundreds or even thousands of different substances that have their origin in the objects that were burned. Scientists say that determining the number of different substances in soot is virtually impossible because of the enormous amount of complexity and the high rate of variables from which soot is produced.

 

Suffice it to say that soot, like smoke, is highly toxic both when inhaled and exposed to the skin.

 

Coping with Smoke and Soot

 

Smoke and soot are a part of life, whether they are generated by a natural forest fire, the smokestack of a chemical factory, soot build-up inside a chimney or the soot that clings to and penetrates everything inside a home in the wake of a fire.

 

That’s why there is an entire industry of professionals who are highly trained to resolve damage issues related to soot and smoke in the aftermath of a fire -- or for routine maintenance, as when a chimney sweep professional removes soot and ash build-up as a preventative measure.

 

Written by Taylor McKnight, Author for Fresh Sweeps

Pharmaceutical Microbiology Resources (http://www.pharmamicroresources.com/)