Monocyte Activation Test for pyrogen testing of biopharmaceutical products

Commonly used acronym: MAT

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Scope of the method

The Method relates to

Human health

The Method is situated in

Regulatory use - Routine production

Regulatory use - GMP process validation

Type of method

In vitro - Ex vivo

This method makes use of

Human derived cells / tissues / organs

Species from which cells/tissues/organs are derived

Human blood

Type of cells/tissues/organs

Peripheral Blood Mononuclear Cells (PBMC)

Specify the type of cells/tissues/organs

Peripheral Blood Mononuclear Cells (PBMC)

Description

Method keywords

PBMC
ELISA
endotoxins and non-endotoxin pyrogens
alternative to rabbit pyrogen test
IL-6
european pharmacopoeia

Scientific area keywords

pyrogenicity
Biopharmaceuticals

Method description

Pharmaceutical products intended for parenteral use must be free from pyrogenic (fever–inducing) contamination. Pyrogens comprise endotoxin from Gram-negative bacteria and non-endotoxin pyrogens (NEP) from Gram-positive bacteria, viruses, and fungi. The longstanding compendial test for pyrogens is the Rabbit Pyrogen Test (RPT) but in 2010 the Monocyte Activation Test (MAT) for pyrogenic and pro-inflammatory contaminants was introduced into the European Pharmacopoeia (Ph. Eur.) as a ‘non-animal’ replacement for the RPT. The developed MAT method was fully validated for GMP purposes according to Ph. Eur. MAT, Quantitative test, Method A to test for pyrogenic and pro-inflammatory substances in therapeutic monoclonal antibodies (mAb). The MAT uses cryo-preserved PBMC with an interleukin-6 (IL-6)-based ELISA readout. The method has been successfully approved by EMA in scope of commercial licensing applications (MAA) for several mAb-based drug products.

Lab equipment

- CO2 incubator;
- Washer;
- ELISA plate reader;
- SoftMax Pro.

Method status

Internally validated
Published in peer reviewed journal

Pros, cons & Future potential

Advantages

- In vitro test which replaces rabbit-based testing;
- (Semi-) Quantitative.

Challenges

- Extensive validation required;
- Requires well-characterized PBMCs.

Modifications

In case the drug product would interfere with an IL-6-based readout, other cytokines such as IL-1 beta may need to be explored and validated as alternative.