EndoLISA^®: A New Endotoxin Detection Method

Endotoxins are of great importance, both in research as well as in the pharmaceutical industry. It is known, that endotoxin contamination can have a dramatic impact on the cultivation of cells and in animal models as well as in humans it induces an immune response that may lead to death. Therefore, monitoring of endotoxin levels is essential.

In many cases, currently used methods for endotoxin detection do not match with the needs of the user, since the sample must be highly diluted, in order to perform the analysis. In addition, currently applied methods for endotoxin testing are using the blood of horseshoe crabs and thereby endanger their existence.

EndoLISA^® is an innovative method for endotoxin detection that completely avoids the use of animal components and renders sample dilution obsolete.

The EndoLISA^® technology uses a phage protein that binds the endotoxin quantitatively to a microwell plate. The solid phase and the binding molecule have been developed to allow the detection of all endotoxin variants with equal affinity. After immobilization of the endotoxin to the solid phase, potentially interfering substances of the sample matrix are removed by a washing step.

The washing step allows the user to work in a wide pH tolerance range (pH 4 to pH 9) and to use samples, which contain a high salt content. EndoLISA^® permits testing samples, which contain concentrations of up to 1 M NaCl, KCl or guanidinium chloride. Furthermore organic solvents, detergents and protease inhibitors are tolerated by EndoLISA^® in much higher amounts than classical methods would do.

Following the washing step, the endotoxin content is quantified by means of recombinant Factor C (rFC) and a fluorescent substrate. The use of recombinant Factor C ensures that the results of EndoLISA^® match with those of the existing assay formats and leads to reliable results.

Each EndoLISA^® kit contains two 96-well microtiter plates. Each plate is divided into 6 separate strips, so that it is not necessary to use the entire plate, when analyzing fewer samples. All needed assay reagents are included in the kit and thus contribute to the overall user-friendly concept of this new endotoxin detection method.

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