Triton X-114液相分离法去除流感单价病毒原液中内毒素的实验研究  

Method of the removal endotoxin from influenza monovalent virus concentrate by triton X-114 liquid phase separation

作　　者：黄秋香[1] 毕湖冰[2] 张新文[2] 高菁霞[2] 蔡玮[2] 刘泽[2] 姚忠萍[2] 姜述德[2] 李映波[2] 

机构地区：[1]云南大学单克隆抗体工程技术中心,昆明650091 [2]中国医学科学院/北京协和医学院,医学生物学研究所,昆明650118

出　　处：《重庆医学》2010年第4期385-387,I0001,共4页Chongqing medicine

基　　金：国家"863"基金资助项目(2006AA02Z409);云南省自然科学基金资助项目(2004C0720M;2006XY29)

摘　　要：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 probe the efficacy of endotoxin in monovalent influenza virus concentrate is removed by Triton X-114 liquid phase separation. Methods Endotoxin was removed by Triton X-114 liquid phase separation,whic^h final concentration were 1% ,2% and 3% separately. The endotoxin content, before and after removal, was detected by tachyplens ameboyto lysate assay. The content of protein was tested by Lowry assay, HA titer by chicken-hemagglutination inhibition test and the HA content was measured by single radial immunodiffusion assay. Results The endotoxin content of monovalent influenza virus concentrate was reduced to less than 0.25EU/mL after the third extraction by Triton X-114,the ratio of reducing was up to 99% ,and no changes in HA tJter,the recovery ratio 05 protein was 85. 11%, and the change of HA content was very little. The best final concentrate al Triton X 114 to remove endotoxin from influenza virus concentrate was 1%. Conclusion Triton X-114 liquid phase separation is an effective method for removing endotoxin from monovalent influenza virus concentrate.

关 键 词：流感单价病毒原液 内毒素 TRITON X-114 

分 类 号：R392[医药卫生—免疫学]