摘要
亚洲鲎血细胞溶解物先经CM-Sepharose CL-6B分级洗脱,再经Dextran-Sepharose CL-6B亲和层析纯化凝固蛋白原。用此法纯化的凝固蛋白原免疫家兔产生抗血清,并用火箭免疫电泳法测定鲎血细胞溶解物与内毒素反应后的凝固蛋白原残余量,根据残余凝固蛋白原量与参加反应内毒素量呈负相关关系,建立了定量测定内毒素的火箭免疫电泳鲎试验法。
Coagulogen from Tachypleus Amoebocyte Lysate(TAL)was isolated and.purified by stepwise gradient elution on CM-Sepharose CL-6B and affinity chromatography on Dextran-Sepharose CL-6B.The chromatographic result for purification of coagulogen was compared with Nakamura's method of gel filtration on Sephadex G-75 and ion-exchange linear gradient elution on CM-Sepharose CL-6B.A monospecific antiserum was obtained by immunizing rabbits with coagulogen purified by affinity chromatography.The antiserum was applied on rocket immunoelectrophoresis to quantify the residue amount of coagulogen after the reaction between TAL and endotoxin,and the amount of endotoxin was consequently determined.TAL Test—Rocket Immunoelectro- phoresis was applied in the diagnosis of endotoxemia in this study.
关键词
凝固蛋白原
内毒素
火箭免疫电泳
Coagulogen
Tachypleus amoebocyte lysate
Rocket immunoelectrophoresis
Endotoxin
