Rapid method for preparing a beta-glucan-specific sensitive fraction from Limulus (Tachypleus tridentatus) amebocyte lysate.
By separating Limulus amebocyte lysate by cation-exchange chromatography with an SP-Toyopearl 650C column, a fraction insensitive to endotoxin, yet specifically sensitive to beta-glucan, was successfully obtained in the unadsorbed portion. This fraction showed beta-glucan dose-dependent clotting enzyme activity, although no sensitivity to endotoxin. This beta-glucan-dependent reaction showed no interference in the presence of endotoxin, with the fraction also showing sensitivity towards various kinds of beta-glucan, i.e. curdlan, pachyman, laminaran and lichenan. The sensitivity towards curdlan was approximately 10(-10) g/ml.
Kitagawa
T
T
Taiyo Central Research Institute, Taiyo Fishery Co., Ltd., Ibaraki, Japan.
Tsuboi
I
I
Kimura
S
S
Sasamoto
Y
Y
D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005936:Glucans / Q000378:metabolism*; D006737:Horseshoe Crabs / Q000166:cytology / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000378:metabolism; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D047071:beta-Glucans
D001427:Bacterial Toxins; D005936:Glucans; D011134:Polysaccharides; D011135:Polysaccharides, Bacterial; D047071:beta-Glucans; C038459:curdlan; C008247:laminaran; C014731:pachyman; C015968:lichenin