We are working with monocyte derived dendritic cells for immunotherapy purposes. As a part of cell culture control we want to test the cells for Endotoxine. We start using the LAL PYROGENT GEL CLOT from LONZA with negative results during standardization of positive controls since July 2014 until today 2015. In november I changed the original kit of 0.03 EU/ml for a new one of 0.06 EU/ml with similar results, and again I changed for a new one 0.125 with similar results. The reproducibility of the test was very week, from one day to the next, the positive controls didn't always form the gel cloth. I usually use water LAL, tubes, tips pyrogenics. I did the test in different laboratories, and I test in incubator at 37ºC, and also in Water Bath. The positive control doesn't work. What can I do with endotoxin gel cloth standarization?
Have you contacted Lonza? Because these kits are designed for human use purposes they are made very stringently, I would be very surprised if it was the kit. I would recommend you go back to the original protocol from Lonza and see if there is a step you missed. The other thing you can do is get a colorimetric endotoxin kit. That kit has you build a standard curve, so you can better find your LOD. Also, as the colorimetric kit starts with very high endotoxin to build the standard curve, if it doesn't work you know there is a problem in how you ran it.
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