Improved bioluminescence-based endotoxin measurement method using a salt-resistant luciferase mutant

Abstract

Endotoxin is a component of the cell wall of gram-negative bacteria and causes fever and shock symptoms upon entering the bloodstream. We previously demonstrated that the bioluminescence-based Limulus amebocyte lysate test is highly sensitive and rapid for measuring endotoxin. However, as the firefly luciferase reaction is inhibited in the presence of sodium chloride, the endotoxin detection method did not meet the validation guidelines under medical dialysis conditions (range of 75-125% of the measured values tested in water). Here, we used a salt-resistant luciferase mutant, which met the criteria for validation of endotoxin measurement.

Keywords: Detection; Dialysis; Endotoxin; Limulus amebocyte lysate test; Luciferase.