Aseptic and Sterile Processing: Control, Compliance and Future Trends

Here is the most important text discussing aseptic and sterile manufacturing to be published in the last decade that looks at both today and tomorrow in regard to these two vital processing procedures.

The Editors realized that there was an urgent imperative for the relevant subjects to be reassessed and represented. To achieve this objective, along with many subject matter experts, they produced a book that is foremost practical. It has been designed for those involved with aseptic and sterile processing to take away many learning points and apply these principles to aseptic and sterile processing within the pharmaceutical and healthcare sectors.

Drawing on experience, they made every effort to incorporate sound science into the practices described, not least to emphasize why new paradigms are required but to provide wide-ranging guidance and offer depth and scope. This is why chapters on human error, risk assessment, depyrogenation, bioburden testing and so on, are extensively covered. It is the aim of the Editors to help readers reassess legacy definitions and historical understandings and move them toward concepts that will help them think in new ways about equipment and processes that will reach the highest standards and evaluate them through science-based risk assessments.

Available from the PDA Bookstore: https://store.pda.org/ProductCatalog/Product.aspx?ID=3850

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

How to Choose and Validate Your Ready-to-Use (RTU) Media

Media that is bought into labs in a ready to go or Ready to Use (RTU) format goes by many different descriptions such as Ready Prepared Media, Pre-Plated Media/Pre-Poured Media (PPM), Ready-to-Rehydrate;  there's no shortage of terms just as there is an almost limitless choice of presentations from broths in bags, ready to rehydrate media in bags and film plates, solid agar in bottles for melting and prepoured plates. But how to choose a supplier that will be best for your lab?

Rapid Microbiology are carrying an exclusive interview with Barbara Gerten, Senior Scientist Traditional Microbiology with Merck KGaA we find out what makes a good media supplier and how to introduce a new RTU media into your lab's schedule.

For details see: https://www.rapidmicrobiology.com/news/how-to-choose-and-validate-your-ready-to-use-rtu-media

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Effective training for keeping cleanrooms clean

Controlled environments required for the manufacture of pharmaceutical products are cleanrooms and they are assigned a class through meeting a set standard for airborne particles. Once a grade is assigned, a series of other physical and microbial parameters need be met: HEPA filtration, control of the air through air changes and pressure differentials; staff wearing appropriate garments; adequate cleaning and disinfection. While such measures can be introduced through the Quality by Design approach and good procedures, to achieve ongoing control requires effective training.

In relation to these important issues, Tim Sandle has written an new article. The introduction reads:

“Traditional training has served the industry well, but it does not always deliver expected outcomes. Moreover, classroom-style training can be expensive and where it is not sufficiently engaging; the message does not always sink in. For these reasons, many pharmaceutical companies are turning to e-learning.”

The link to the article is: https://www.cleanroomtechnology.com/news/article_page/Effective_training_for_keeping_cleanrooms_clean/148134

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Recording device for cell history

Different "events" such as infections by viruses, as well as the exposure to environmental toxins or other forms of stress, change the activity of genes thereby leaving molecular traces inside the cell. These changes happen mainly at the level of messenger RNA (mRNA). These are molecules that encode genetic information when genes become activated and read, a process known as transcription. Researchers can accurately investigate the activity of a gene by measuring the mRNA molecules present in a cell. However, the traces of gene transcription disappear rapidly: mRNA is highly instable, and cells often degrade it after a short time.

ETH researcher Randall Platt and his colleagues in the Department of Biosystems Science and Engineering have now developed a molecular recording system that writes transcriptional events into DNA where they can be permanently stored and later accessed to by sequencing.

To create their "recording device," Platt's doctoral students Florian Schmidt and Mariia Cherepkova employed the CRISPR-Cas system. CRIPSR-Cas is an adaptive immune system in bacteria and archaea. The system functions like an immunological memory device by recording genetic information about pathogens infecting the cell. This genetic information is recorded in a specific stretch of DNA known as a CRISPR array -- a process called acquisition.

CRISPR arrays are capable of storing short sequences of DNA, known as 'spacers', originating from a pathogen. Spacers are separated from each other by short identical DNA sequence called direct repeats, just like pearls on a string.

The researchers worked with the gut bacterium Escherichia coli, introducing the genes for the CRISPR-Cas system from a different bacterial species. One of those Cas genes is fused to a reverse transcriptase, an enzyme that uses an RNA molecule to produce DNA encoding the same information -- in other words, it transcribes RNA back into DNA.

The Escherichia coli cells supplied with the foreign genes for this CRISPR-Cas were able to produce a protein complex that binds short mRNA molecules. The reverse transcriptase translates these RNA spacers into DNA, containing the same information as the original RNA, and subsequently storing them in the CRISPR array. This process can occur multiple times such that new spacers are added to the CRISPR array in reverse chronological order, so the most recently acquired piece of DNA is always first.

In principle, this makes it possible to record any number spacers within a CRISPR array. Since DNA is very stable, the information recorded in them is stored for a long time and is also passed on from one generation of bacteria to the next.

"Our system is a biological data logger. It records the genetic response of bacteria to external influences and enables us to access that information even after many bacterial generations" says Florian Schmidt, the lead author of the study, which was recently published in the journal Nature.

ETH Professor Randall Platt says, "Researchers have been working on creating forms of synthetic cellular memory for a long time, but we are the first to develop one that can record information about the expression of each gene in a cell over time." The researchers have spent over two years working on this system.

Until now, researchers were limited to measuring mRNA at only a single snapshot in time. Taking these snapshots generally means destroying the cell, extracting its mRNA, and then quantifying them. In contrast, the new CRISPR-Cas RNA recording system records the history of cell, allowing researchers to effectively access the entire cellular log book rather than just a single point in time.

As part of their study, the ETH researchers recorded the reaction of E. coli bacteria equipped with the data logger to the herbicide paraquat. This substance provokes changes in mRNA transcription within the cells, and the scientists could read out this response from the CRISPR arrays even days after the herbicide exposure. Without the data logger, any molecular traces of the bacteria's contact with the herbicide would have long since been broken down and the information lost.

Biological data loggers like this, in addition to being interesting for research purposes, could also conceivable be used as a kind of sensor, to measure environmental toxins such as the herbicide, or in diagnostics. The present study intriguingly demonstrates the feasibility of such an approach, however practical applications are still a long way off. Randall Platt's research team in Basel is already working on transferring the system to other cell types and paving the way for its effective use as a diagnostic tools.

See:

Florian Schmidt, Mariia Y. Cherepkova, Randall J. Platt. Transcriptional recording by CRISPR spacer acquisition from RNA. Nature, 2018; DOI: 10.1038/s41586-018-0569-1

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Happy New Year

Happy New Year to all readers of Pharmaceutical Microbiology - let's look forward to lots of exciting microbiology news for 2019.

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Good pour plate practices

The Microbiologics blog has a useful back-to-basics article on good pour plate practices. Included in the article is the following advice:

1. Keep the molten agar in the water bath for no more than three to four hours. Don’t pour the agar until it has cooled to <50°C (preferably 44°C to 46°C).

2. Don’t re-melt the agar. Agar should only be melted one time.

3. Use phosphate buffer pH 7.2 if necessary to dilute the suspension.

4. Decrease the risk of contamination by pouring plates in a laminar-airflow cabinet. When pouring multiple plates, flame the mouth of the flask before moving on to the next plate to reduce the risk of contamination.

5. Fill plates according regulators’ recommendations. The U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) recommends filling plates with 12 ml to 15 ml of agar. The United States Pharmacopeia (USP) recommends a fill of 15 ml to 20 ml of agar.

6. Some microorganism species, such as obligate aerobes, may recover better on spread plates than pour plates. When growing these strains, it is recommended to verify counts with a spread plate.

7. Incubate most bacterial species for 48 to 72 hours. Note: Incubate Candida albicans and Aspergillus brasiliensis for three to five days.

8. Count small microorganism colonies, such as Pseudomonas aeruginosa, with the aid of an illuminated colony counter or magnifying glass.

9. Keep in mind recovery will be lower on selective agar. If selective agar is used, test non-selective agar in parallel using the same microorganism suspension. A higher CFU concentration for the selective agar may be necessary.

10. Don’t be surprised if the value obtained when performing tests differs from the mean assay value. Note: Microbiologics uses non-selective Tryptic Soy Agar when testing most microorganisms.

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Happy holidays!

I'd like to wish all readers of Pharmaceutical Microbiology all the best wishes for the holiday season and thank you for supporting this website.

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Pharmig Guide to Bacterial Identification

Pharmig’s latest publication is a guide to bacterial identification. The guide discusses why identification is important and what needs to be identified, answering the often-challenging questions of ‘what’, ‘when’ and ‘how often’?

There are several text books on identification. These, however, err towards the clinical. Texts on identification approaches for pharmaceutical microbiology are not common and guidance on understanding the appropriate level of identification is difficult to obtain.

Microbial identification represents an important part of the microbiology function. This includes screening products for objectionable organisms, profiling the environmental microbiota, and investigating out-of-limits events with a view to assigning a probable point of origin. In deciding what and when (and subsequently to what level) to identify, and by the way of which methods, requires an identification strategy. This is a document each microbiology laboratory should develop.

Many parts of pharmaceutical microbiology are outlined in compendia or in guidance documents issued by regulators; included within these are the importance of bioburden assessments of intermediate and finished products, and the need to monitor the environment using standard environmental monitoring methods. What is less clear is expectation with regards to microbial identification. For identification, there are established and emerging methods, based around the microbial phenotype or genotype, yet the choice between systems is not straightforward and the selection depends, in part, on what needs to be identified. Deciding which types of samples to identify; what level of identification is appropriate (morphology, genus, or species); and what can be done with the collected information needs careful thought.

Written by Dr. Anna Lovatt (GSK) and Dr. Tim Sandle (BPL), the guide discusses different methods for phenotypic and genotypic identification, and the latest rapid methods. Troubleshooting sections and case notes are included with each section. The guide comes with a foreword from Andrew Hopkins of the MHRA.

The reference is:

Lovatt, A. and Sandle, T. (2018) Guide to Bacterial Identification, Pharmig, Stanstead Abbotts, UK

For details contact Pharmig

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Use of Liquids and/or Soft Foods as Vehicles for Drug Administration

FDA has issued new draft guidance: “Use of Liquids and/or Soft Foods as Vehicles for Drug

Administration: General Considerations for Selection and In Vitro Methods for Product Quality Assessments Guidance for Industry.”

The introduction reads: “This guidance applies to orally administered drug products and provides recommendations to sponsors who will use or recommend use of liquids3 18 and/or soft foods as vehicles for drug administration in investigational new drug applications (INDs), new drug applications (NDAs), Biologics License Applications (BLAs), as applicable, and in supplements to these applications. This guidance addresses the approaches recommended for suitability determination of vehicles intended for use with specific drug products by providing the following…Considerations for selection of liquids and/or soft foods as vehicles…Standardized in vitro methodology and data recommendations for drug product quality assessments to qualify vehicle(s) for drug product administration… Recommendations to communicate acceptable (qualified) vehicles in drug product labeling. If certain foods are found unacceptable, they should also be included in the labelling.

See: https://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM614401.pdf

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Pharmig Annual Conference 2018

Pharmigs 26th Annual Conference Programme Interactive 2018 Final (1) by Tim Sandle on Scribd

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

GMP in Practice: Regulatory Expectations for the Pharmaceutical Industry

James Vesper and Tim Sandle have a new book – ‘GMP in Practice: Regulatory Expectations for the Pharmaceutical Industry’, Fifth Edition, Revised and Expanded.

The Long-Awaited Revision and Update of GMP in Practice is Here!

Have you ever asked yourself, "Where in the Good Manufacturing Practices (GMPs) does it say I have to do _______?" If so, look no further than PDA's GMP in Practice: Regulatory Expectations for the Pharmaceutical Industry, fifth edition, Revised and Expanded.

As companies strive to harmonize global requirements for quality systems, the 5th edition of this text provides an overview of the 34 essential global cGMP requirements that are typically included in a modern pharmaceutical quality system, including data integrity and how they have evolved. Explore risk-related questions, delve into several expectations for each quality system element encompasses, and review real-world examples from cGMP regulations from the US FDA, Health Canada, the European Union, the World Health Organization, and the International Conference on Harmonization (ICH).

If you're looking for an enhanced understanding of GMP in practice, this text is a must-have for your reference collection.

See: https://store.pda.org/ProductCatalog/Product.aspx?ID=4511

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Avoiding environmental monitoring ‘false negatives’

In addition to selecting the right culture media, the use of an appropriate neutraliser is important in relation to surface, and some personnel, monitoring. Neutralisers are required to overcome any residues left by disinfectants, as can be found on cleanroom surfaces or on the gloved hands of personnel. The use of a neutraliser within the culture media formulation is also necessary to overcome residues from antimicrobial compounds so that a false negative is avoided.

The use of a neutraliser is recommended in the biocontamination control standard ISO 14698;3 and, outside of pharmaceuticals, the cosmetics microbiological test standard ISO 21149 contains some useful advice on neutraliser selection.

Tim Sandle has written a new paper on the topic of neutralisers for culture media. The introduction reads:

“The selection of an appropriate neutraliser is not straightforward. The neutraliser must be non-toxic to the microorganisms expected to be recovered; be able to stop residual disinfectant activity; and, in use. This latter requirement often proves the most challenging. This article examines the most common neutralisers used; some of the problems associated with their selection; and the complexities around using the most appropriate neutralisers in the culture media most commonly used in the environmental monitoring programme.”

The reference is:

Sandle, T. (2018) Avoiding environmental monitoring ‘false negatives’: overcoming disinfectant residues with culture media neutralisers, European Pharmaceutical Review, 23 (4): 18-21

The article can be accessed here: https://www.europeanpharmaceuticalreview.com/wp-content/uploads/epr418-EnvMonitoring-IDF.pdf

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Working with Biohazards

Working with human pathogens or biohazards poses serious risks, not only for employees, but for the public and communities as well. Infectious agents such as microorganisms, viruses, recombinant or synthetic nucleic acid molecules, and biological toxins present a potential for severe or lethal disease, adverse health effects, or contamination. Any unplanned exposure or release has the potential to cause extensive harm or damage to people, the environment, and society.

Vince McLeod has written an interesting article for Laboratory Manager. Here is an extract:

“The foundation for safe handling and research with infectious/biohazardous agents is an effective exposure control plan (ECP). This article discusses the basic elements of a comprehensive exposure control plan, what each element should address, and advice for successful implementation.

The ECP is essentially a biohazard safety manual developed to address the unique conditions of the current research, facility design, and personnel operations necessary to carry out the laboratory’s mission. One excellent free reference is the CDC’s Biosafety in Microbiological and Biomedical Laboratories1 (BMBL), which contains comprehensive information on biological risk assessment and summary statements on many common infectious agents.”

To access the article, see: https://www.labmanager.com/lab-health-and-safety/2018/05/working-with-biohazards

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

NIBSC experts will be discussing how control materials are relevant

Join us and @IBMScience at our upcoming webinar where NIBSC experts will be discussing how control materials are relevant. Book now: https://t.co/GJKm6FqYdm pic.twitter.com/BNCozPkSfu

— NIBSC_MHRA (@NIBSC_MHRA) October 29, 2018

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology

Advanced Technologies are Tackling the Global Lyme Epidemic

More than 300,000 new cases of Lyme disease, the most common tick-borne illness in the United States, are diagnosed each year. In a new interview with CMRubinWorld, Dr. Brian Fallon, Director of the Lyme and Tick-Borne Diseases Research Center at the Columbia University Irving Medical Center reveals that despite the challenges to find a cure for this complex, debilitating disease, precision medicine and biotechnology are accelerating the discovery of new tools with which doctors will be able to diagnose it and treat patients.

In an authoritative new book, Columbia University Medical Center physicians Brian Fallon and Jennifer Sotsky explain why there is much cause for optimism. “Through rapid genetic sequencing, scientists can identify many different strains of Borrelia burgdorferi (causative agent of Lyme disease) as well as new tick-borne microbial infections, such as Borrelia miyamotoi, Borrelia mayonii, and the Heartland virus,” says Fallon. The discovery of these new microbes inside ticks has significantly helped researchers since it “provides a starting point for the study of pathogenesis, vaccine development, and treatment.” Fallon notes that researchers have also been able to screen thousands of drugs to determine which have the ability to destroy Borrelia.

Read the full article here

Brian Fallon, MD, MPH is the Director of the Lyme and Tick-Borne Diseases Research Center at the Columbia University Irving Medical Center and the author with Jennifer Sotsky of Conquering Lyme Disease: Science Bridges the Great Divide, published in 2018 by Columbia University Press.

CMRubinWorld’s award-winning series, The Global Search for Education, brings together distinguished thought leaders in education and innovation from around the world to explore the key learning issues faced by most nations. The series has become a highly visible platform for global discourse on 21st century learning, offering a diverse range of innovative ideas which are presented by the series founder, C. M. Rubin, together with the world’s leading thinkers.

For more information on CMRubinWorld

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology