Adventitious Agent Testing For MMV: How Much Do We Really Know?

Minute virus of mice (MMV) has contaminated biotechnological processes in the past and specific MMV testing is therefore recommended, if the production cell line is known to be permissive for this virus. Testing is widely done using cell-culture based adventitious virus assays, yet MMV strains may differ in their in vitro cell tropism. Here, I present growth characteristics of different MMV strains on A9 and 324K cells and identified significant differences in susceptibility of these widely used indicator cell lines to infection by different strains of MMV, which has implications for MMV detectability during routine testing of biotechnology process harvests [1]. An MMV specific PCR was evaluated as a more encompassing method and was shown as suitable replacement for cell culture-based detection of the different MMV strains, with the additional benefit that detection is more rapid and does also cover other rodent parvoviruses that might contaminate biotechnological processes. Although no MMV contamination event of human-derived cell lines has been reported in the past, biotechnological processes that are based on these also need to consider MMV-specific testing, as we showed e.g. HEK293, a human-derived cell line commonly used in biopharmaceutical manufacturing as susceptible to productive MMV infection.

[1] Farcet et al., Biotechnology & Bioengineering 2024; 121:131-138; DOI: 10.1002/bit.28573

Presented by Maria Farcet, Head of Cell Culture, Virus Models & Serology in the Global Pathogen Safety / Global Quality team at Takeda

Maria Farcet is Head of Cell Culture, Virus Models & Serology in the Global Pathogen Safety / Global Quality team of Takeda in Vienna, Austria. In her 15+ year career in this field she gained profound expertise in virology and virus safety margins of biological medicinal products.  She currently oversees the establishment and operation of virus test systems to enable virus reduction studies, virus testing and virus neutralization assays, under GMP/GLP.

Advancing Viral Safety in Biologics: Comprehensive Virus Detection with iDTECT® Transcriptome NGS-Based Assay

The emergence of broad molecular methods raised some gaps in traditional viral safety testing of biologics. This presentation highlights the potential of iDTECT® Transcriptome, a Next-Generation Sequencing (NGS) based assay designed for the viral safety testing of cell substrates. By targeting viral RNA transcripts expressed during replication, the transcriptomic approach enables comprehensive detection of both RNA and DNA viruses across a variety of species, including rodent, human, simian, bovine and porcine cells. The results show high sensitivity, with a detection limit of 0.1 RNA copies per cell, achieving virus detection levels that are comparable to or exceeding conventional methods. The NGS transcriptomic approach is effective in identifying both close and distant viral strains, making it a more comprehensive, sensitive, and cost-effective alternative to existing methods.

Amina Cherif Louazani, PhD, Viral Safety Manager at PathoQuest

Amina Cherif Louazani (PharmD, PhD), Viral Safety Manager at PathoQuest, has a strong background in virology and bioinformatics. She holds a PhD in Infectious Diseases from Aix-Marseille University, France, and specializes in NGS data analysis and protocol development for viral transcriptomes. Amina has published several peer-reviewed articles and held various research and clinical roles. At PathoQuest, she leverages her expertise to provide expert reviews of viral safety assessments based on the iDTECT® platform of NGS assays.  

Viral safety testing of human cell lines used for biopharmaceutical manufacturing.

Testing of cell banks is required by the regulatory authorities. Different approaches can be used to meet these expectations. The webinar will focus on the regulatory landscape concerning cell bank testing (with a focus on safety concerns), as well as on the traditional methods and qPCR approaches, that are commonly used. It will further highlight the advantages and disadvantages of these methods, and the gaps that come along with targeted testing.  

Dr. Denise Teber, Scientific Expert of the PCR department at Charles River Laboratories

Dr. Denise Teber is the Scientific Expert of the PCR department at Charles River, Erkrath, Germany. She is responsible for the establishment of new qPCR and ddPCR assays for the detection of pathogens or process-related biological impurities, as well as method validation of platform assays or client-specific requests. She has a biology degree from the University of Münster and a Master’s degree from the University of Cologne. During an external master thesis at the German Aerospace Center (DLR), she investigated the influence of ionizing radiation on cell cycle progression to predict its harmful impact on astronauts on the International Space Station – ISS. During her PhD and post-doctoral work at the Max Planck Institute for Molecular Biomedicine in Münster, she used lentiviral and adenoviral vectors to induce putative mutations causing venous malformations in isolated primary mouse endothelial cells and analyzed their effects by qPCR, co-immunoprecipitation and western blot as well as intravital microscopy.

Sponsored by PathoQuest and Charles River Laboratories

About PathoQuest

PathoQuest is a global CRO providing Next Generation Sequencing (NGS)-based Quality Control testing through its iDTECT® assay platform. This platform enables faster, safer and animal free testing for the release of biotherapies to patients. We offer GMP-validated assays for viral safety and genetic characterization of raw materials, cell and gene therapies, monoclonal antibodies and vaccines. Our laboratories in Paris, France, and Wayne, PA, USA, provide global and reliable support to accelerate the availability of biologics. Learn more: https://pathoquest.com/

About Charles River Laboratories

No matter where you are in your drug development journey, Charles River can provide you with a wide range of products, models, services, and guidance that are tailored to your specific needs. Our biologic and manufacturing services help you navigate seamlessly throughout the development cycle with a wide variety of comprehensive CGMP testing and manufacturing solutions including monoclonal antibodies/recombinant proteins, viral vectors, vaccines, and cell therapies. With over 40,000 tests run annually at our global biologics testing facilities and 200+ products supported; you can count on our expertise to ensure your therapy is safe, effective, and able to pass regulatory scrutiny.