Endotoxin and Pyrogen Testing - Challenges for Biotechnological and Biopharmaceutical Products

In the context of the current development of new or alternative microbiological methods and the attempt to reduce the use of laboratory animals throughout Europe, the monocyte activation test in particular has gained in importance. It has been part of the European Pharmacopoeia for quite some time. Now it will be published in an own chapter.

The monocyte activation test (MAT) is used to detect or quantify the effect of substances that activate human monocytes or monocytic cells to release endogenous mediators such as pro-inflammatory cytokines, e.g. tumour necrosis factor alpha (TNFα), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6). These cytokines are important in the pathogenesis of fever. The MAT thus detects the presence of pyrogens in a sample. After a product-specific validation, the MAT can be used as a replacement for the rabbit pyrogen test.

Pharmaceuticals or biopharmaceuticals containing non-endotoxin pyrogenic or pro-inflammatory impurities often show very steep or non-linear dose-response curves compared to endotoxin dose-response curves. Therefore, preparations which contain or may contain non-endotoxic contaminants usually have to be tested at a defined minimum dilution in a specific dilution range. In general, three MAT methods are differentiated:

• Quantitative test 
• Semiquantitative test 
• Limit test

Currently, a number of test kits are available on the market, but the application in the laboratory is still new for many laboratories.

The ECA offers a special laboratory practical training "Monocyte Activation Test" on 13/14 February in Bernried, Germany, where the different kits will be presented and their correct use is shown and performed by the participants themselves. Experts from the regulatory authorities, from the laboratory and from the test manufacturers will introduce the special features of the method, the possible applications and the practical implementation. The number of participants is limited due to practical laboratory work in small groups. This course can also be booked in combination with the "Low Endotoxin Recovery Laboratory Course" on 11/12 February.