人肠道病毒71型VP4多抗的制备及其应用

发布时间：2018-05-27 00:01

本文选题：肠道病毒型 + VP　；参考：《中国生物制品学杂志》2016年03期

【摘要】：目的制备人肠道病毒71型(enterovirus 71,EV71)VP4多抗,并鉴定EV71空心颗粒(empty particle,EP)和实心颗粒(full particle,FP)。方法构建重组表达质粒p GEX-6p-1-VP4,诱导表达并纯化GST-VP4融合蛋白,免疫家兔制备多抗。ELISA法检测抗体效价,免疫荧光试验(immunofluorescence assay,IFA)和Western blot法鉴定抗体特异性。应用制备的多抗经Western blot法区分EV71 EP与FP。结果质粒p GEX-6p-1-VP4经双酶切及测序鉴定证明构建正确,纯化后的GST-VP4融合蛋白相对分子质量约32 000,以包涵体形式存在。免疫家兔后可获得效价达10~(10)的多抗。兔抗EV71 VP4多抗可识别EV71原核表达及天然病毒颗粒中的VP0、VP4。结论抗EV71 VP4多抗可应用于EV71 EP及FP的鉴定,为EV71的基础研究及疫苗研发提供了新的工具及方法。
[Abstract]:Objective to prepare a polyclonal antibody against human enterovirus 71 (EV71) and to identify EV71 hollow particles, empty particles and solid particles. Methods Recombinant expression plasmid pGEX-6p-1-VP4 was constructed, and the GST-VP4 fusion protein was induced and purified. The antibody titers were detected by immunofluorescence assay (IFA) and Western blot method. The prepared polyclonal antibody was used to distinguish EV71 EP from EV71 by Western blot. Results the plasmid p GEX-6p-1-VP4 was correctly constructed by double enzyme digestion and sequencing. The relative molecular weight of purified GST-VP4 fusion protein was about 32 000, which existed in the form of inclusion body. After the rabbits were immunized, the titer of the polyclonal antibody was 10 ~ (10). Rabbit anti EV71 VP4 polyclonal antibody can recognize EV71 prokaryotic expression and VP0 VP4 in natural virus particles. Conclusion the anti EV71 VP4 polyclonal antibody can be used in the identification of EV71 EP and FP, which provides a new tool and method for the basic research and vaccine development of EV71.
【作者单位】：武汉生物制品研究所有限责任公司病毒性疫苗研究一室;
【分类号】：R392

【相似文献】