Selective assay for endotoxin using poly(epsilon-lysine)-immobilized Cellufine and Limulus amoebocyte lysate (LAL).
We developed a selective endotoxin (lipopolysaccharide; LPS) assay using poly(epsilon-lysine)-immobilized cellulose beads (PL-Cellufine) and Limulus amoebocyte lysate (LAL). First, LPS was selectively adsorbed on the beads in a solution containing various LAL-inhibiting or LAL-enhancing compounds (e.g., amino acids, enzymes) and the LPS adsorbed on the beads was separated from the compounds by centrifugation. Second, the LPS adsorbed on the beads directly reacted with the LAL reagent, and the LPS concentration was determined by a turbidimetric time assay. The accuracy of the adsorption method with PL-Cellufine was high compared with that of a common solution method. Apparent recovery of LPS from compound solution was 88-120%.
Sakata
Masayo
M
Department of Applied Chemistry & Biochemistry, Graduate School of Science and Technology, Kumamoto University, Kumamoto, Japan. msakata@gpo.kumamoto-u.ac.jp
Fukuma
Yuriko
Y
Todokoro
Masami
M
Kunitake
Masashi
M
D000327:Adsorption; D000818:Animals; D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D004731:Endotoxins / Q000032:analysis*; D007447:Invertebrate Hormones / Q000737:chemistry*; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D008863:Microspheres; D009391:Nephelometry and Turbidimetry; D011107:Polylysine / Q000737:chemistry*; D012107:Research Design
D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D004731:Endotoxins; D007447:Invertebrate Hormones; D008070:Lipopolysaccharides; D008565:Membrane Proteins; C034705:antilipopolysaccharide factor (Limulus); C025357:endotoxin binding proteins; D011107:Polylysine