本文選題：腸道病毒型 + VP�。� 參考：《中國生物制品學(xué)雜志》2016年03期

【摘要】：目的制備人腸道病毒71型(enterovirus 71,EV71)VP4多抗,并鑒定EV71空心顆粒(empty particle,EP)和實(shí)心顆粒(full particle,FP)。方法構(gòu)建重組表達(dá)質(zhì)粒p GEX-6p-1-VP4,誘導(dǎo)表達(dá)并純化GST-VP4融合蛋白,免疫家兔制備多抗。ELISA法檢測抗體效價(jià),免疫熒光試驗(yàn)(immunofluorescence assay,IFA)和Western blot法鑒定抗體特異性。應(yīng)用制備的多抗經(jīng)Western blot法區(qū)分EV71 EP與FP。結(jié)果質(zhì)粒p GEX-6p-1-VP4經(jīng)雙酶切及測序鑒定證明構(gòu)建正確,純化后的GST-VP4融合蛋白相對(duì)分子質(zhì)量約32 000,以包涵體形式存在。免疫家兔后可獲得效價(jià)達(dá)10~(10)的多抗。兔抗EV71 VP4多抗可識(shí)別EV71原核表達(dá)及天然病毒顆粒中的VP0、VP4。結(jié)論抗EV71 VP4多抗可應(yīng)用于EV71 EP及FP的鑒定,為EV71的基礎(chǔ)研究及疫苗研發(fā)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.
【作者單位】： 武漢生物制品研究所有限責(zé)任公司病毒性疫苗研究一室;
【分類號(hào)】：R392

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本文編號(hào)：1939495