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目的探讨Triton X-114液相分离法去除流感单价病毒原液中内毒素的效果。方法用终浓度为1、2、3%Triton X-114抽提、液相分离法去除流感单价病毒原液中的内毒素;用鲎试剂凝胶法检测去除前后内毒素含量、Lowry法测定蛋白质含量、红细胞凝集试验检测血凝效价、单相免疫扩散法测定血凝素含量等。结果通过3轮抽提,Triton X-114能够将单价病毒原液中内毒素水平降至低于0.25EU/mL,去除率达到99.9%,蛋白质回收率为85.11%,血凝效价保持不变,血凝素含量几乎无变化。以1%Triton X-114去除流感单价病毒原液中内毒素效果最好。结论Triton X-114液相分离法可有效去除流感病毒原液中的内毒素。
Objective To investigate the effect of Triton X-114 liquid phase separation on the removal of endotoxin from influenza virulent stock solution. Methods Triton X-114 was used to extract the endotoxin in the solution of influenza monovalent virus by liquid phase separation. The content of endotoxin in the monovalent virions was detected by gel electrophoresis. The content of protein was determined by Lowry method , Hemagglutination test hemagglutination test, single-phase immunodiffusion assay hemagglutinin and so on. Results After 3 rounds of extraction, Triton X-114 could reduce the level of endotoxin in the monovalent virus solution to less than 0.25EU / mL, the removal rate reached 99.9%, the protein recovery rate was 85.11%, the blood coagulation titer remained unchanged, Hemagglutinin content almost no change. The best removal of endotoxin in the influenza monovalent virus stock solution with 1% Triton X-114. Conclusion Triton X-114 liquid phase separation method can effectively remove the endotoxin in the influenza virus stock solution.