Detection of lipopolysaccharide in the picogram range of tissue culture media by a kinetic chromogenic Limulus amebocyte lysate assay.
A kinetic chromogenic Limulus amebocyte lysate assay for detection of lipopolysaccharide in the picogram range is described. The kinetic Limulus assay was used to detect lipopolysaccharide in a tissue culture media containing interfering low-molecular-weight substances. The interfering substances were removed by ultrafiltration. The recovery of lipopolysaccharide from the ultrafiltrated tissue culture media was 61.6 +/- 21.4%. The kinetic Limulus assay was compared with the traditional end-point chromogenic Limulus assay. The results in the two assays showed a good correlation. The detection limit was 1 pg/ml in the chromogenic assay and 0.25 pg/ml in the kinetic assay.
Royal Danish School of Pharmacy, Department of Biological Sciences, Copenhagen.
M R
MR
Pedersen
E W
EW
Hansen
J D
JD
Christensen
D000595:Amino Acid Sequence; D000814:Aniline Compounds; D000818:Animals; D002863:Chromogenic Compounds; D003470:Culture Media; D046508:Culture Techniques; D006737:Horseshoe Crabs; D007700:Kinetics; D008033:Limulus Test / Q000379:methods*; D008070:Lipopolysaccharides / Q000032:analysis*; D008832:Microchemistry; D008969:Molecular Sequence Data; D012680:Sensitivity and Specificity
D000814:Aniline Compounds; D002863:Chromogenic Compounds; D003470:Culture Media; D008070:Lipopolysaccharides; C008625:nitroaniline