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Endotoxins and their Relevance in R&D ApplicationsLPS Detection & Removal Methods

endotoxins are detected via mediators (e.g. II 1 , II 6, TNF-a). Thereby mainly monocytes or leukocytes like RAW264.7, MonoMac 6 or THP 111 are used.

The in-vitro pyrogenic test (IPT) is a patented procedure to detect pyrogenic determination and is based on humane whole blood. It is an alternative for the rabbit test since it detects also positive bacteria and others biological pyrogenics (such as exotoxines, lipoteichoic acid, parasitical constituents) which cannot be verified in the LAL.

They are also methods which use the 3-Hydroxy fatty acids of the lipopolysaccharides for the endotoxine detection via gas chromatography/mass spectrometry.

The LPS concentration is indicated in EU/ml or EU/mg. The unit EU (endotoxin unit) describes the biological activity of LPS. 1 EU is generally converted to 100 pg LPS.

A Gram-negative bacteria contains approximately 10-15 g of LPS, 1 EU therefore can be generated by only 105 bacteria.


LPS Removal
Removal of endotoxin is one of the most difficult downstream processes during protein purification. Many commercial available products are unable to remove endotoxin satisfactorily, or require time consuming incubation steps. In many cases, complete endotoxin removal is only achieved with massive substrate loss. Reduction or removal of endotoxin to less than 1 ng/mg (10 EU/mg) is a very difficult task. Only methods with highest endotoxin removal capacity combined with excellent recovery rates of the target substance are reasonable and acceptable. To meet exactly these most challenging requirements, Profos AG has developed an endotoxin trap for lab- and pilot-scale: EndoTrap.

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