Endotoxin assay by bioluminescence using mutant firefly luciferase.
The Limulus reaction is an application of the defense mechanism of horseshoe crab for endotoxin detection. Endotoxin is a component of the cell wall in the outer membrane of gram-negative bacteria, and causes fever or shock when it enters the human blood stream. For endotoxin detection, gel formation or turbidity of the coagulation factor chromogen or fluorescence-modified peptide is used. However, these conventional methods have problems with regard to their measurement time or sensitivity. We recently obtained a mutant firefly luciferase that has a luminescence intensity over 10-fold higher than that of the wild type. Therefore, we developed a new endotoxin detection method that combines the Limulus reaction and bioluminescence using mutant luciferase. The new method detects 0.0005EU/ml of endotoxin within 15min.
Noda
Kenichi
K
Research Institute for Nanodevice and Bio Systems, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8530, Japan. noda@hiroshima-u.ac.jp
Goto
Hitoshi
H
Murakami
Yuji
Y
Ahmed
Abo Bakr F
AB
Kuroda
Akio
A
D000818:Animals; D001681:Biological Assay; D004731:Endotoxins / Q000032:analysis*; D008033:Limulus Test / Q000379:methods*; D049409:Luciferases, Firefly / Q000235:genetics* / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*
D004731:Endotoxins; D049409:Luciferases, Firefly