Validation and Implementation of the Monocyte Activation Test as an Alternative Method to the Rabbit Pyrogen Test  

The journey of implementing the MAT as an alternative to the RPT

Implementing the Monocyte Activation Test (MAT) is more than a technical switch; it is a strategic organizational journey. This presentation outlines the roadmap for transitioning from the Rabbit Pyrogen Test (RPT) to an animal-free alternative, starting with the identification of the business need and the definition of clear project goals. We will explore the “implementation journey” by defining the scope, resource allocation, and timeline required to establish a robust pyrogen testing capability.

A critical component of this roadmap is navigating the complex regulatory landscape. While the European Pharmacopoeia (Ph. Eur.) accepts MAT as a compendial method to replace RPT, the United States Pharmacopeia (USP) currently views it as an alternative method requiring full method validation. We will detail the specific expectations of global Health Authorities, providing a guide on how to align internal business objectives with external regulatory compliance. This presentation provides a step-by-step framework for managing the change to a scientifically advanced, ethical, and compliant pyrogen testing strategy.

Presented by Jonas Van Den Berg, Subject Matter Expert, Pyrogen Testing and Alternative Microbiological Methods at Roche

Jonas van den Berg joined Roche in 2017, where his expertise is currently focused on the practical implementation of new Quality Control technologies. He serves as a Subject Matter Expert for pyrogen testing and alternative microbiological methods. Over his career, he has evaluated a number of innovative microbiological technologies for their suitability in regulated environments. Jonas van den Berg is currently the Project Manager for the implementation of the Monocyte Activation Test. This project not only introduces a reliable testing method but is also a key contribution to the company’s animal welfare strategy, successfully validating the MAT as an alternative to the traditional Rabbit Pyrogen Test. His role leverages his extensive practical experience in pharmaceutical operations, analytical QC, and complex qualification and validation projects. Before joining Roche, Jonas van den Berg worked as a consultant, specializing in microbiological method development for the pharmaceutical industry. He earned his Ph.D. in microbiology and biochemistry from the University of Groningen, the Netherlands.

Validation Strategy and Practical Implementation of the MAT

The Monocyte Activation Test (MAT) is an in vitro assay based on human blood cells capable of detecting both endotoxins and non-endotoxin pyrogens (NEPs), offering a scientifically and ethically superior alternative to the Rabbit Pyrogen Test (RPT). To ensure successful implementation across product types, this presentation outlines a two-tiered validation strategy that aligns with both European and U.S. regulatory expectations. The first tier consists of a generic, product-independent validation following the semi-quantitative MAT method according to the Ph. Eur., as well as ICH Q2 and USP <1225>. Validation parameters included Range and Linearity, Limit of Detection, Accuracy, Specificity, Precision, and a comprehensive assessment of Robustness. Robustness was evaluated under routine-relevant conditions such as stimulation time, IL-6 read-out timing, freeze–thaw stability, and lot-to-lot comparability. Additionally, an equivalency study comparing the RPT and the MAT demonstrated that MAT performance is at least non-inferior, if not superior. This structured approach provides a GMP-compliant and transferable framework for laboratories seeking to implement the MAT reliably and efficiently.

Presented by Luisa Burgmaier, Project Leader, Endotoxin and Pyrogen Testing Group at Microcoat Biotechnologie

Luisa Burgmaier holds an M.Sc. in Biotechnology and joined Microcoat Biotechnologie GmbH as part of her doctoral research. Her work centers on endotoxin and pyrogen testing, with a strong focus on improving analytical reliability and understanding the underlying mechanisms that affect detection. As part of her research, she investigates endotoxin masking phenomena and evaluates factors that influence detection and recovery in complex biological matrices.

Within Microcoat’s Endotoxin and Pyrogen Testing Group, Luisa serves as a Project Leader, overseeing both research-driven initiatives and customer-focused projects. In this role, she also manages customer projects involving the Monocyte Activation Test (MAT), supporting method development, workflow optimization, and the implementation of reliable pyrogen testing strategies. She collaborates closely with partners to develop robust analytical approaches and ensure high-quality testing solutions tailored to their needs.

Followed by a live Question and Answer session