Biomedical applications of Limulus
amebocyte lysate.
To find an easy and efficient way to wipe off the bioburden, we examined three approaches (methods A, B, and C) with different flow rates and flushing durations, using common reagents (75% ethanol, water, and acetic acid), and checked the results using limulus
amebocyte lysate (LAL) clotting test.
Endotoxin levels in the samples were determined using the key quality characteristics limulus
amebocyte lysate.
Plasma endotoxin concentration was determined by kinetic turbidimetric assay (Yokota et al., 1989), using reagent kit bought from Xiamen tachypleus
amebocyte lysate Co.
In order to analyze concentrations of endotoxins and peptidoglycans, frozen filters were warmed up to room temperature and then subsequently eluted with 10 ml of Limulus
amebocyte lysate (LAL) reagent water (Lonza, Ltd., Basel, Switzerland) with 0.05% Tween 20 (Sigma-Aldrich, Ltd., Poznau, Poland) on a platform shaker Tetramax 1000 (Heidolph, Schwabach, Germany) for 15 min, and centrifuged with a force of 1000xg for another 15 min.
An aliquot (0.1 ml) of stock solution processed above was added to 0.1 ml Limulus
amebocyte lysate (LAL) reagent, and the kinetic turbidity of the mixture was measured using a tube reader (Zhanjiang A and C Biological Ltd., China).
(9) The basis of the Fungitell assay relies on BG's ability to activate the limulus
amebocyte lysate clotting cascade present in the blood of Limulus polyphemus, the North American horseshoe crab.
Their blue blood contains cells called Limulus
amebocyte lysate (LAL) that, when used properly, can detect bacteria.
Limulus
amebocyte lysate (LAL) assay and in vitro macrophage activation assay
The endotoxin content of these preparations was negligible as determined by the limulus
amebocyte lysate assay (Endos, USA).