The PyroGene Recombinant Factor C Assay offers an animal-free alternative for endotoxin testing. For a long time substances have been tested for bacterial endotoxins using Limulus Amoebocyte Lysate (LAL), which is extracted from horseshoe crabs.
The recombinant form of Factor C, which is the first component in the endotoxin-activated coagulation cascade of horseshoe crabs, is activated by endotoxin binding and the active enzyme then cleaves a synthetic substrate, resulting in the formation of a fluorogenic compound. This method described here works through a single enzymatic step compared to the multiple-step enzymatic process necessary for LAL assays, with the same reliability.
The method will be included in the European Pharmacopoeia from 2021.
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