Tim Sandle

The University of Manchester · School of Pharmacy and Pharmaceutical Sciences

Cleanroom microflora are of importance for microbiologists and quality control personnel in order to assess changes in trends. Shifts in the types of microflora may indicate deviations from the "norm" such as resistant strains or problems with cleaning practices. Given the few published studies of the typical microflora, this paper uniquely reviews...

Aims: To examine for psychrophilic or psychrotolerant micro-organisms in pharmaceutical cold rooms (in relation to numbers, incidents and species) and to determine, where such micro-organisms are present, whether standard microbiological environmental monitoring regimes require modification. This is presented as a case study. Methods and results:...

Cleanroom wipes, saturated with disinfectant, are commonly used within pharmaceutical grade cleanrooms as part of contamination control programmes. Whilst standards are in place for the testing of wipes for particle shedding, there is a limited data relating to the particle generation from wipes when they are used in practical conditions. This pape...

Whilst the pharmaceutical microbiologist now faces a somewhat bewildering array of different test systems, once the selection has been made it is important, from a GMP perspective, that the system is validated. For this, both the United States Pharmacopeia and European Pharmacopeia offer some guidance. In general, the two approaches share more simi...

The aim of this study was to describe the incidence of contamination of pharmaceutical products by melanized fungi and to consider control measures in relation to bioburden and cleanrooms. This study reviews and analyses pharmaceutical product recalls and offers incidence rates of fungal detection from a typical cleanrooms. The recalls include some...

Maintaining compliance for the sterility test, demonstrating the validated state, enabling testing consistency, and showing traceability of consumables-each an important component of laboratory data integrity-has been made easier through the introduction of Merck's M-Trace ® system. This article looks at the importance of sterility test compliance...

Fungal contamination in pharmaceutical products represents a potential hazard for two reasons. First, it may cause product spoilage; the metabolic versatility of fungi is such that any formulation ingredient may undergo chemical modification in the presence of a contaminating organism. Spoilage not only affects the therapeutic properties of a produ...

Ideas for extending automation to the endotoxin test has primarily been approached through machinery, to improve assay precision, and through other forms of automation to improve data integrity in terms of sample traceability (including data transfer to laboratory information management systems) and to enable the assessment of data patterns and tre...

AI/ML is predicted to be the top technology investment for 60 percent of life sciences companies over the next two years, according to a recent survey from the Pistoia Alliance. In this chapter, advances of AI within the field of microbiology are considered, focusing on technologies that are enhancing the governance of contamination control and w...

The new Annex 1 places even more emphasis on the risks of cross-contamination within aseptic production areas. Not surprisingly, it offers several specifications about the transfer of materials: the use of material pass-boxes with integrated biodecontamination systems, as well as airflow trolleys, becomes essential.

All microorganisms require some amount of water in order to reproduce. Microorganisms take up water by moving it through their cell membranes. This is why, even with all the necessary growth factors, water is proportionately the largest ingredient within culture media. Water is required for metabolic activity and as a biophysical factor. The amount...

There are different factors which influence water system contamination. Good design features are concerned with generation processes, keeping organisms out of water systems and having in-built controls to reduce the possibility of survival. One factor that influences the potential for microbial survival is the presence of organic material – signifi...

Microbiological culture media is either a solid, liquid or semi-solid composite designed to support the growth of microorganisms. It is represented as the commonly seen solid agar in a dish (the Petri dish) or liquid broth in a bottle. Liquid media are used for encouraging growth or for continuously maintaining reproduction, as with pure batch cult...

Advanced Therapy Medicinal Products (ATMPs) are gene therapy medicinal products, somatic cell therapy medicinal products and tissue engineered products (1). These therapies, without hyperbole, offer ground-breaking new opportunities for the treatment of diseases and injuries. They are particularly important for severe, untreatable or chronic diseas...

While rapid microbiological methods have advanced, most microbiology laboratory tests remain reliant upon assessing microbial growth on agar plates. Discrepancies with plate counting, together with the limitations of human vision, have led to regulatory concerns to the extent that the U.S. FDA Q&A document on data integrity recommends “double plate...

The modern microbiology laboratory is starting to look very different from the laboratory of 10 years ago, and in the next few years, the microbiology laboratory is set to appear different from the laboratory of today. The work will still involve assessing microbial numbers, determining susceptibility, and identifying organisms, and there will cont...

There are the fungal organisms that pose the greatest risk to human health worldwide. In all, fungi form the World Health Organisation (WHO) priority pathogens list (FPPL). One of the aims is to help to direct global health efforts to research into methods to tackle fungal pathogens, considering that many of the fungi and the diseases they causes r...

An important component of pharmaceutical products and medical device research and development is a focus on the users of the device. Developing an effective device or medication can be easily undone if it is not used correctly, and errors made by patients or clinicians are often a direct result of design limitations.1 Weaknesses can be overcome by...

On the basis of Food and Drug Administration (FDA) of United States of America recall data from 2012 to 2022, heterotrophic microorganisms have caused the majority of microbial contamination incidents reported in non-sterile pharmaceutical products. Some of the products impacted are liquids, tablets, capsules, oils, drops, creams and emulsions. The...

Assessing microbiological data and undertaking investigations into the origins of contamination, establishing root causes, and setting appropriate preventative actions constitutes a core part of the contamination control strategy. Despite the importance of this process, regulatory findings frequently cite poor quality microbial investigations. Thi...

The study aimed to manage and to analyse the results of the laboratory tests, routinallypractice for hepatitis C diagnosis, using blood tests. Statistical analysis of this study results, was performed using the laboratory informatic system. The results of the study are substantial and intricate reffering to comparision between two methods concretll...

For reworking, a pharmaceutical manufacturer needs to have in place a procedure that includes details on retesting and reevaluation criteria for the nonconforming product after rework. Reworking should be associated with a deviation record (for the original reason to consider rework) and executed under change control. As well as standard product-re...

For several decades isolators, as separative devices, have been commonplace in pharmaceutical and healthcare products manufacturing. Although isolators presented a significant step forward on the contamination control continuum, isolators, if not appropriately operated, can be vulnerable to contamination ingress. The primary mechanism for ingress i...

While these are well-established facts, the ability of M. luteus to survive for prolonged periods within the cleanroom environment is less commonly understood. Excluding endospores, for the common vegetative bacteria, M. luteus displays a level of robustness for prolonged survival within the cleanroom if it is not directly eliminated using a disinf...

A major challenge with antimicrobial drug development is the necessity to test millions of drug compounds in the hope of finding one that will kill a given microbial pathogen. As part of the search for new antimicrobial compounds designed to be effective against emerging drug-resistant bacteria, laboratories are using robotic chemical-synthesizing...

Nanomedicine has advanced considerably in the past five years (as per the OECD nanotechnology R&D intensity measure). Nanomedicine is a broad term, defined as using nanoscale materials, including biocompatible nanoparticles and nanorobots for disease diagnosis, drug delivery, physiological sensing, or for actuation purposes. The innovations in nano...

For effective cleaning validation, appropriate limits need to be set to identify risks of cross-contamination from equipment surfaces, especially where there is shared equipment used across different process streams. To do so requires an understanding of which indirect surfaces pose the greatest likelihood of contamination transfer together with an...

The guide has been completely revised and re-written to provide you with a roadmap to regulatory compliance for cleaning & disinfection. The new text will walk you through the steps needed to design, validate, and implement an effective cleaning and disinfection programme.

For both sterile and non-sterile manufacturing, bioburden control as evidenced by microbial counts is especially important in relation to the quality of the finished product and as an indicator of process control. Samples are drawn from intermediate product at defined stages (ideally based on risk assessment) and these allow for the microbial level...

Rapid microbial tests should be risk-based so the stakeholder can select the preferred technology for their intended use and balance user requirement specifications including time to result, specificity, limit of detection, sample size, and product attributes. Many rapid methods produce signals not based on a colony forming unit (CFU), and that som...

Several studies have demonstrated the effectiveness of antimicrobial polymeric flooring (such as the special flooring manufacturing by Dycem). These studies have focused on particulate removal and reductions in microbial levels. What has been less-well studied is the suitability of this type of flooring for the reduction in fungal counts. This arti...

Cleaning validation receives a great deal of attention within the biopharmaceutical industry, not least because of the risks of product adulteration and hence patient harm from improperly cleaned surfaces (notwithstanding additional concerns such as operator protection). Traditionally, cleaning validation efforts focus on direct product-contact sur...

For the past two decades, nanotechnology has promised a revolution in medicine, especially in creating “smart” ways to deliver medicines to specific targets for more effective treatments. Scientists are working on various types of nanoparticles, especially to diagnose and treat cancer. This article considers some of the key innovations within the f...

Anyone working in drug manufacturing knows Dr. Tim Sandle. A microbiologist, author, and science journalist, he is one of the leading experts and figurehead in the field. For this reason, we asked Dr. Sandle for his opinion on the effects of the new ANNEX 1 in the strategies that, with due differences, the entire industry will have to implement: f...

This paper identifies the risks of contamination by viruses in the manufacture of biopharmaceutical products in cleanrooms. Different types of viruses are described with non-enclosed viruses being more resistant to disinfectants. Care should be taken as many common disinfectants are only suitable for bacterial or fungal contamination. A table gives...

The study aimed to manage and to analyse the results of the laboratory tests, routinally practice for hepatitis C diagnosis, using blood tests. Comparison of ELISA method results (Enzyme-Linked Immunosorbent Assay) and chemiluminescence methods results. Statistical analysis of this study results, was performed using the laboratory informatic system...

For aseptically filled products, as well as for many terminally filled products, the sterility test is a mandatory product release test. It is, however, statistically poor at detecting anything other than gross contamination (this limitation has been addressed in a number of studies (1)). This limitation relates to the few numbers of articles teste...

There are different elements that contribute to good aseptic technique within the cleanroom and the laboratory. One such element is the donning of gloves (1), handling items appropriately, and keeping gloves regularly disinfected (2). Glove disinfection is an essential step for bacteriological control, although how successful control is maintained...

For isolators to be used effectively, they need to be operating to EU GMP Grade A / ISO 14644 class 5. This is achieved through the barrier concept, the maintenance of a positive pressure, the supply of HEPA filtered air and its extraction, and by subjecting the isolator to an automated decontamination cycle. Isolators are most commonly sanitised u...

Chronic liver diseases stimulate a degree of hepatocyte injury. This previously mentioned modifications, alters the known liver architecture and finally ends in cirrhosis. Liver pathology as cirrhosis develops after a long period of pathological alterations. The management of this liver pathology, is centred on the treatment of the causes and compl...

The pathology of childhood occurring with increased frequency is tonsillitis. The infection affects both sexes equally, without any association with environmental factors. The symptomatology attracts attention and requires treatment by a specialized medical team. The severity of the symptoms often leads to surgery to remove these formations via a t...

Gene therapy products are required to be safe, stable, of low toxicity, and of a particular cell type specificity. In some cases, viral vectors are engineered to infect a wide range of cell types; in other cases, the viral vector is modified (pseudotyped) to target a specific kind of cell.1 An important step in the development process is assembling...

With dual-incubation regimes, a key variation in practice concerns the starting temperature and order of incubation. Moving from higher to lower temperatures can inhibit the growth of fungal contamination, not only because fungal growth is optimized at low temperatures, but also because high temperatures could damage the lytic cellular enzymes that...

Graphically representing data is more than just creating pretty pictures; proper data visualization can improve the communication of sometimes abstract information to make data accessible, understandable, and usable. Patterns and insights arise during the review process — whether data are intended for internal or external publication or for inclusi...

Cleaning and disinfection are necessary protective and reactive steps to address microbial deposition on surfaces within pharmaceutical cleanrooms. An appropriately designed cleaning and disinfection program plays a central role in a contamination control strategy. There is considerable literature on the importance of cleaning and with disinfectant...

An important objective of hygienic design relating to the pharmaceutical and healthcare sectors is with the avoidance of product contamination by microorganisms, particles and chemicals. When considering cleaning validation, microbial and chemical deposits and their removal require different approaches and they are influenced by different factors (...

Improving the time-to-result and doing so with greater accuracy has become a topic of concern for the microbiology laboratory as much as it has for other economic sectors. Achieving the result early may improve the chances of patient survival or it may help a firm to reduce inventory hold times. There is a wide spectrum of rapid microbiological me...

Late in 2022, the FDA authorized a new vaccine including an adjuvant, available under emergency use authorization to prevent COVID-19 in individuals 12 years of age and older. The important word is “adjuvant,” the use of which is sometimes open to misinterpretation, especially with traditional adjuvants made from aluminium salts. An adjuvant is a...

Scientific data is concerned with measuring and hence data, whether that is qualitative or quantitative. In pharmaceutical microbiology, this could be a number of cells or colony forming units, a series of growth or no growth results; and incidences of microorganisms. Gathering such data allows for trending and enables control to be achieved. With...

Analytical techniques for validating cleaning methods must evolve to match the pace of an increasingly complex manufacturing landscape for new therapeutics. Within the bio/pharma industry, there is a certain expectation for companies to maintain required levels of cleanliness within their facilities to ensure products that are manufactured are at...

For a safe batch release, a pyrogen test should be preferred to avoid missing out on non-endotoxin pyrogens in the final drug product. Monocyte Activation Test, based on human model and with a high sensitivity is a good option for batch release.

The objective of the guidance is to outline the important safety aspects and data verifications to support efficacy claims for studies of multiple versions of a cellular or gene therapy product in an early-phase clinical trial for a single disease within a single clinical trial. These studies are not intended to provide primary evidence of effectiv...

Toward the end of 2022, the FDA issued guidance on the validation of biological assays, titled M10 Bioanalytical Method Validation and Study Sample Analysis.1 The guidance was originally issued in draft form in 2019; a little over three years was required to develop the final guidance document, and it was approved by ICH in May 2022. Hence, the FDA...

A contamination control strategy (CCS) is a system that considers all the integral elements of pharmaceutical product manufacturing. A CCS should cover: • Microbial contamination • Cleaning and disinfection • Sterility assurance • Facility design • Chemical and particle contamination

Medicines are chemicals that are used to cure, slow, or prevent disease. Some medicines are injected into the bloodstream, so production facilities must make sure those medicines do not contain microorganisms, toxins, or particles. If they did, the medicines could make people very ill and even cause death, which is the opposite of what medicines ar...

The disposal of pharmaceuticals and the waste generated from pharmaceutical production has an environmental impact, particularly in waterways. The extent of this varies by geography and in relation to the extent of local regulations and the willingness to comply with these regulations. Environmental concerns foremost relate to the detection of trac...

Sections 4.33 to 4.36 of the 2022 edition of EU GMP Annex 1 focus on cleaning and disinfection. In this GMP blog, we look at cleaning and disinfection techniques. For effective cleaning, the application technique is important. Wiping strategy includes the applied pressure force, wiped surface area, the geometry of the mechanical action, the number...

One of the most commonly used agents for disinfection are quaternary ammonium compounds. Discovered to be antimicrobial agents over one hundred years ago, products made from the compounds entered into widespread use after World War II. As well as being the active ingredient of a vast array of disinfectants, the compounds are used in fabric softener...

The US Food and Drug Administration (FDA) has issued a new white paper on the subject of Quality Management Maturity (QMM). This outlines a framework by which the Agency will assess pharmaceutical manufacturers using a ratings scale. This involves looking for evidence that signals an organisation has moved beyond simply complying with current good...

The new version of EU GMP Annex 1 was issued on 22nd August 2022 1 , addressing the manufacture of sterile products. This is a legally binding part of EU GMP (as per Article 47 of Directive 2001/83/EC on the Community code relating to medicinal products for human use of the European Union). It also applies to the UK, as part of the Medicines Act of...

Microbial air samplers are used to collect a predetermined volume of air and operate in a way to remove any microorganisms and to capture the microorganisms onto an agar-based growth medium. Once the sample has been collected and the medium incubated, the results are typically expressed in colony forming units per cubic metre (cfu/m3). Active air...

A lean laboratory should deliver results as efficiently as possible — in terms of cost, time, and quality compliance. The objectives of a lean laboratory program can include: to improve definition, structure, and control of laboratory processes; to increase the consistency and predictability of laboratory performance; to produce a detailed understa...

The ultimate path that precision medicines will take is unclear, although it is already apparent that diseases that hitherto were classed as 'untreatable' can potentially be treated (and, in a few cases, are being treated). These conditions are genetic diseases and certain cancers. While scientific thinking has advanced and technologies have follow...

Purification is a necessary phase in pharmaceutical drug manufacturing in order to eliminate unwanted materials that can be hazardous or which will compromise drug efficacy. This is of particular concern with protein-based drug products, where impurities include closely related non-functional proteins. To achieve the desired purification, attention...

In pharmaceutical plants and through distributing loops, pipes can leak (as with pharmaceutical water systems). Leaks can also occur in relation to vessels and through tubing or connectors designed for the product path(as part of single use disposable technology set-ups). Such leaks pose a risk in terms of microbial intrusion. One of the myths is t...

Viruses threaten to spoil biotechnological products at nearly every manufacturing stage. Viral contamination can affect raw materials, cell culture processes, bioreactor contamination, and downstream processing. ICH has released a new draft guideline titled Viral Safety Evaluation of Biotechnology Products Derived From Cell Lines of Human or Animal...

Cleaning validation receives a great deal of attention within the biopharmaceutical industry, not least because of the risks of product adulteration and hence patient harm from improperly cleaned surfaces (notwithstanding additional concerns such as operator protection). Traditionally, cleaning validation efforts focus on direct product-contact sur...

A major theme is antimicrobial agents (and the role played by pharmaceutical microbiologists in developing and delivering these agents) and the growth in microbial resistance. In the last two decades, the rate at which bacteria are becoming resistant to current antibiotic treatments has substantially increased. This trend is threatening the ability...

Is anaerobic environmental monitoring necessary? For obligate anaerobes the case for doing so is low. For facultative anaerobes, where aseptically filled parenteral products are involved, the case is far higher. Any such consideration should be based on a quality risk assessment taking into account the interactions between people and exposed produc...

The main reason for aseptic sampling failing is inadequate training. A training scheme will need to cover avoidance of poor sampling habits and means to avoid the shedding of microflora from the body. Poor sampling habits can include the use of non-sterile equipment or the improper use of the equipment.

Messenger RNA (mRNA) vaccines made global headlines in 2021 in the fight against the SARS-CoV-2 virus responsible for COVID-19 (through the Pfizer-BioNTech and the Moderna vaccines). The principle behind these vaccines is to use engineered mRNA to provide instructions to cells to signal the immune system to create antibodies against specific viral...

The regular application of disinfectants represents a key measure for controlling the numbers and types of microorganisms present within a healthcare or pharmaceutical facility. There is an array of different types of disinfectants, each with a different method of microbial kill and with efficacy being influenced by different external factors. Impo...

To assess medicines for safety and efficacy, it is important that clinical trials are representative of biological sex. Too often, there is an underrepresentation of women in clinical trial subject populations. This leads to erroneous data since women differ in presentation, clinical manifestations, and outcomes in comparison to men.2 For example,...

Many items entering cleanrooms, especially aseptic processing facilities, are required to be sterile. Sterilisation by an ionising radiation source is the most common way of achieving sterility and is used for cleanroom garments as well as for plastic items intended for single use, such as sampling containers, aseptic connectors, biocontainer bags,...

In this article, pharmaceutical microbiologist and contamination control expert Tim Sandle presents three microbial contamination investigation case studies, highlighting the key lessons for pharmaceutical microbiologists to take away and the underlying importance of identifying the root cause of microbial data deviations. One of the main activiti...

Bacterial endonucleases, cleave DNA molecules in strictly specific areas, generating fragments of varying lengths, called restriction fragments (FR). Restriction enzymes (ER) have as an essential feature the specificity of their action. This may explain that an ER exerts its endonuclease activity only in the presence of a certain nucleotide sequenc...

Advances in biotechnology have led to a new generation of vaccine treatments entering development and trial. These innovations include the use of genetically modified organisms and vaccines based on mRNA, where human cells are programmed to develop antibodies against specific diseases. While these vaccines appear new to the general public, research...

The realization of medicines based on mRNA technology is a relatively recent breakthrough, with several products under trial. Moreover, the COVID-19 vaccine success is spurring a further stream of medicines to come. An mRNA-based medicine is a different concept; it does not fall into small molecule-based pharmaceuticals or within conventional biolo...