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Sustainable testing

Parenteral pharmaceutical products are required to be free of pyrogens that may induce life-threatening systemic reactions in a patient. Globally harmonized quality control tests to determine pyrogens are the Rabbit Pyrogen Test (RPT), established in 1912, and the Bacterial Endotoxin Test (BET), commercialized since 1977. However, as the world is increasingly concerned with the ethics of using experimental animals for pyrogen testing pharma manufacturers seek to establish in-vitro pyrogen test methods eliminating animals from QC testing. Moreover, there is growing need to test for non-endotoxin pyrogens that pose a risk to human health and to overcome the limitations faced by traditional animal-based pyrogen tests with the complex formulations and manufacturing processes used for biologics, including vaccines or cell and gene therapies. 

With the  PyroCell® Monocyte Activation Test Rapid System and the PyroCell® MAT Human Serum Rapid System you can now bring rapid, sensitive and reliable detection of endotoxins and non-endotoxin pyrogens (NEP) to a test plate, saving experimental rabbits while ensuring safe medicines.

The next generation of pyrogen testing kits

Lonza launches the PeliKine Human IL-6 Rapid ELISA Kit allowing detection of IL-6 cytokines in just 2 hours - now included in all PyroCell® MAT Rapid System Kits.

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Monocyte activation test - Assay principle

With Monocyte Activation Tests, the response of the human innate immune system to pyrogens is measured. The assay uses human monocytes, the key cells of innate immunity, that respond to the presence of endotoxins or non-endotoxin pyrogens by secreting pro-inflammatory cytokines such as interleukin-6 (IL-6) linked to the fever reaction in humans. With PyroCell® Monocyte Activation Tests, the released cytokines are quantitated using a human IL-6 ELISA assay.


PyroCell® Monocyte Activation Test Rapid Systems are optimized and validated reagent kits for the sensitive detection of pyrogens in pharmaceutical preparations. The PyroCell® MAT Rapid System consists of a cryopreserved peripheral blood mononuclear cell (PBMC) preparation pooled from four human blood donations (pMAT cells), and an optimized, FBS-based Culture Medium Supplement. It has been validated with the Pelikine Human IL-6 ELISA Rapid Kit, ensuring robust and reliable results.

The PyroCell® Monocyte Activation Test Human Serum Rapid System (PyroCell® MAT HS Rapid System) uses the same PBMC preparation and the human IL-6 ELISA detection system but human serum as culture medium supplement. For Monocyte Activation Tests, the use of human serum provides a more sustainable reagent formulation and moreover, has been demonstrated as advantageous for pyrogen testing of blood-based products. On day one of the experiment, the cryopreserved pMAT cells are thawed and incubated with the test substance in a cell culture step (18-24 hours). On the following day, the cell culture supernatants are harvested, and the IL-6 cytokine released by human monocytes is detected with the human IL-6 ELISA assay and measured in an absorbance reader. Using reference endotoxin dilutions, the OD values are finally converted into Endotoxin Equivalents (EE/mL). 

Monocyte activation test - Applications

Early detection of pyrogens in a pharmaceutical preparation is critical for quality control programs and an essential product safety measure. Pyrogens may originate from microorganisms such as bacteria and fungi and also from viruses, genome components or other organic particles from the environment. Pyrogens initiate inflammatory reactions when injected into a patient. With the evolution of manufacturing technologies that deliver for example, vaccines, bioprocessed proteins or cell and gene therapy products, new contamination risks may arise in production. Though the Monocyte Activation Test principle was established decades ago, its adoption for life-saving biologics has recently accelerated. 

PyroCell® Monocyte Activation Test Systems reflect decades of experience in developing and manufacturing specialized MAT components thus providing the perfect solution for life-saving drugs that are unethical to test in experimental rabbits or cannot be tested with standard bacterial endotoxin tests. 

Pyrogen Test Method Comparison

Qualified cells

  • Ethical cell sourcing - blood donations specifically for the Monocyte Activation Test
  • Validated process – ensure endotoxin-free preparations in a validated time frame 
  • Naïve cells - demonstrated with quality certificates for critical parameters 

Cryopreserved donor pools

  • Available on demand — no cell isolations prior to each experiment
  • Large lot size prepared with equal cell quantities of 4 blood donations 
  • Proven robustness and reliability

IL-6 as cytokine read-out

  • Fully secreted into the medium
  • Stable in (frozen) supernatant
  • Ensure highly sensitive detection

Regulatory information MAT

Compliance to regulations requires QC testing of all injectable pharmaceuticals and medical devices for the presence of pyrogens whose effects may range from fever, shock, and in some instances, death. Globally harmonized tests to detect pyrogenic substances are the animal based rabbit pyrogen test (RPT), as well as the bacterial endotoxin test (BET).

The world’s increasing concerns with ethics of using experimental animals, and efforts to protect natural resources led regulatory agencies and pharmaceutical companies to acknowledge in vitro test systems such as the monocyte activation test (MAT) and the recombinant factor C test (rFC) that are minimizing such dependencies. Likewise, the European Pharmacopeia has adopted the MAT in 2009 (Ph. Eur. chapter 2.6.30) and recently, has put an end on the RPT for the year 2026. Other leading pharmacopeia such as the United States Pharmacopeia are describing the MAT as a suitable alternative test method to the RPT.

Other advantages of the MAT include the stimulation of a human response (RPT: mammal response), introduction of experimental controls and cost savings. Therefore, the MAT is closing the gap between patient safety and other regulatory requirements.

Suitability of Pyrogen Test Methods

Benefits of PyroCell® Monocyte Activation Test

Sensitive, robust, reliable

  • Patient safety – detection of the full range of pyrogens in one test
  • No experimental rabbits – acknowledged by regulators to replace the rabbit pyrogen test
  • Human-specific – mimics the innate human response to pyrogens
  • Flexible – 2 validated system kits meeting product-specific and sustainability demands
  • Standardized – robust application and assay controls meet high sensitivity for reliable results 

Road to sustainability
future-proofing QC lab pps

Webinar

Webinar attendees will learn how to increase efficiencies in the QC lab by incorporating sustainable solutions for testing, automation and digitalization.

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Sustainable solutions: Lonza testing

Video
Optimize your workflow and ensure patient safety with our sustainable pyrogen and endotoxin testing solutions. Learn about products we offer and their benefits in this short video. 
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How to swiftly adopt
the MAT

White paper

Our new white paper features an interview where Sanquin’s Dr. Marijke Molenaar-de Backer, Senior Scientist MAT Services, shares important insights into the sustainable pyrogen test.

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