狂犬病毒核蛋白（NP）的纯化及其免疫原性的研究

对重组痘苗毒和重杆状病毒表达的狂犬病毒NP及原代地鼠肾细胞培养的狂犬病毒核衣壳蛋白（RNP）、先经Sepharose CL 4B分子筛柱初步提纯，再经抗狂犬病毒NP McAB 2C12-S-epharose 4B亲和层析柱纯化分离，经ELISA、SDS－PAGE电泳和Western-Blot分析证实，获得了高纯度和免疫反应性的NP和RNP。以相同剂量的纯化蛋白免疫小鼠，RNP和两种重组NP均可诱生特异的抗NP抗体，三种蛋白间无明显差异；狂犬病毒CVS株攻击保护实验结果显示，三种蛋白免疫的小鼠存活率约为50％；两种重组NP的免疫反应性和免疫原性与天然狂犬病毒RNP相似。

Study on Purification and Immunogenicity of Rabies Virus Nucleoprotein

In order to establish the optimum condition for purification of the nucleoprotein(NP) of rabies virus by immunoaffinity chromatography, the efficient and non denaturative eluents(Mg el) was obtained by using ELISA elution model; furthermore, it didn't damage the activity of NP. Two kind of NPs, expressed by recombinant vaccinia virus (rVac N) and recombinant baculovirus (BRN), were purified by a Sepharose CL 4B column and a 2C12 Sepharose 4B column. By Western blot and SDS PAGE, high purity and good antigenical intact NPs were identified. The purified ribonucleoprotein (RNP) of rabies virus 5aG strain was also obtained. After immunized with NP and RNP, mice developed a strong anti nucleoprotein response and were protected against a lethal challenge of rabies virus CVS strain. There were not difference been observed among the mice immunized with different purified protein. These data indicate that the NPs are antigenical and immunogenical comparable to the authentic rabies RNP and therefore present a potential source of an effective ,safe and economical subunit vaccine.