PEG-6000纯化流感病毒鸡胚尿囊液的研究

发布时间：2018-04-25 19:01

本文选题：流感病毒 + 实验室技术和方法　；参考：《国际检验医学杂志》2014年01期

【摘要】：目的探讨PEG沉淀法纯化鸡胚培养流感病毒尿囊液的工艺,对PEG-6000的浓度、沉淀时间、离心力进行优化,为流感病毒抗原和抗体制备提供了便捷的手段。方法向流感病毒鸡胚尿囊液添加PEG后进行高速离心收集沉淀,检测纯化病毒液效价、病毒形态,以及卵清蛋白含量。结果 PEG浓度为4%,沉淀时间2h,离心力8 000g条件下,病毒能完全沉淀,杂蛋白卵清蛋白含量低,病毒颗粒完整,浓缩液中病毒血凝效价为1∶10 240。以纯化病毒免疫山羊3次后,血清抗体滴度为1∶1 024且无交叉。结论本研究初步证明优化后的PEG沉淀法对鸡胚培养流感病毒尿囊液中的病毒浓缩、纯化达到了很好的效果。
[Abstract]:Objective to study the purification process of chicken embryo culture influenza virus allantoic fluid by PEG precipitation method, optimize the concentration, precipitation time and centrifugal force of PEG-6000, and provide a convenient method for the preparation of influenza virus antigen and antibody. Methods PEG was added to the chicken embryo allantoic fluid of influenza virus. The purified virus was collected and precipitated by high speed centrifugation. The titer of purified virus, the morphology of the virus and the content of ovalbumin were detected. Results when the concentration of PEG was 4 and the precipitation time was 2 h, the centrifugal force was 8 000 g, the virus could precipitate completely, the content of hybrid protein ovalbumin was low, the virus particles were intact, and the hemagglutination titer of virus in concentrated solution was 1:10 240. After three times of immunization with purified virus, the titer of serum antibody was 1:1 024 and there was no cross-crossing. Conclusion the optimized PEG precipitation method has a good effect on virus concentration and purification in chicken embryo culture influenza virus allantoic fluid.
【作者单位】：中国医学科学院/北京协和医学院/医学生物学研究所/云南省重大传染病疫苗研发重点实验室;
【基金】：国家国际合作项目基金(2011DFR30420) 云南省自然科学基金(2010CD134)
【分类号】：R373.13

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