抗狂犬病免疫核糖核酸的分离纯化及部分理化性质和生物学活性初探  

Purification and part of physico-chemical characterization as well as biological activity for immuno-globulin ribonucleic acid of anti-rabies

作　　者：杨琳[1] 闫素志[1] 张金岩[1] 柳春红[1] 邱野[1] 李彤[1] 徐颖鑫[1] 夏天瑶[1] 闫肃[1] 

机构地区：[1]长春生物制品研究所,130062

出　　处：《中华微生物学和免疫学杂志》2010年第2期140-143,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的 探索一种抗狂犬病免疫核糖核酸（iRNA）的制备方法 ,为狂犬病病毒暴露后的免疫治疗开辟途径.方法 用狂犬病病毒免疫马,从抗体阳性血清的马匹中分离肝脏,依次采用十二烷基磺酸钠、苯酚、三氯甲烷、乙二醇单甲醚、十六烷基三甲基溴化铵、乙醇等提取总RNA.结果 提纯品经理化性质检测,得到的iRNA占肝脏总重的0.15%,其中DNA含量为2.86%,蛋白质含量为1.26%.最大吸收峰位于258 nm;A258/A280为2.0;RNA鉴别试验呈阳性反应.用高效液相色谱法测定,相对分子质量为13.7×10^3;增色效应50.67%.生物学活性测定结果 显示,白细胞黏附抑制率为41.73%.动物保护率为50%,生命延长率为31.62%.结论 实验提取的抗狂犬病iRNA,经用国家颁发的iRNA质控标准对照检测,结果 相同.为狂犬病治疗药物的研究奠定了实验室基础.Objective To explore the preparation of specific immune RMA（iRNA） on anti-rabies and further study immunotherapy of rabies virus exposure. Methods Horses were immunized with the rabi-es virus and their livers were isolated from the horse of antiserum, from which total RNA was extracted and purified by sodium lauryisulfonate, phenol, chloroform, ethyiene glycol monomethyl ether, cetyltrimethyam-moniumbromide and alcohol. Results Pure preparation physico-chemical characterization was analyzed, and it＇s weight was 0.15% of weight of liver. The RNA contained 2.86% DNA and 1.16% protein. The iRNA with a maximum UV absorbance at 258 nm and A258/280 about 2.0. The test of RNA was positive, which had a relative molecular mas of 13.7×10^3 by high performance liquid chromatography（HPLC）, and its hy-perchromic effect was 50.67%. The vesults of biological activity was showed that the rate of leucocyte adher-ence inhibition（LAI） was 41.73%, The protective rate was 50% and prolonging the life was 31.62%. Conclusion The results obtained with the practical value were identical and provide a basis on medicines of anti-rabies.

关 键 词：抗狂犬病 免疫核糖核酸 纯化 理化性质 生物学活性 

分 类 号：R512.99[医药卫生—内科学]