Large expression and application of the measles virus neutralization antigen H and F protein using the virus engineering technique.

为了对麻疹病毒重要的保护性抗原血凝素蛋白（H蛋白）进行三维结构分析，制备了2种表达H蛋白的重组杆状病毒。除H基因的胞质结构域外，还产生了连接流感病毒A/日本株（H3亚型）HA蛋白信号序列的重组杆状病毒，使得H蛋白可以从感染的昆虫细胞分泌到培养基中。第二种病毒产生重组杆状病毒，其将因子Xa的切割排列引入H蛋白的细胞膜附着蛋白。采用亲和层析、离子交换层析和特异性抗体凝胶过滤层析等方法对H蛋白进行纯化，以大量回收这两种H蛋白。随着纯化阶段的结束，通过H蛋白与麻疹病毒单克隆抗体的反应性降低。那个H pro ...更多信息 与单克隆抗体反应的蛋白仅显示中和抗体，仅存在于电泳和凝胶过滤色谱识别澄清的大聚集体的分级分离中。并且，在任一蛋白中也不能识别绿色猴红细胞的血凝活性。从上述结果来看，分离纯化重组杆状病毒蛋白用于结构分析似乎是不合适的。因此，从病毒粒子中分离纯化蛋白质对于进行病毒的三维结构分析是重要的，并且，我们已经在明胶颗粒中致敏了麻疹病毒，并建立了新的麻疹抗体检测方法。本次应用该方法，建立了麻疹IgM抗体检测方法。世卫组织的麻疹根除计划采用了这种方法，因为这种方法方便，并且将在非洲和南美洲等发展中国家进行检查。

2 kinds of recombination baculoviruses that manifested the H protein were produced in order to carry out three-dimensional structure analysis of the important hemagglutinin protein (H protein) as a protective antigen of measles virus. Except for cytoplasm domain of the H gene, recombination baculovirus which connected the HA protein signal sequence of the influenza virus A/Japan strain (H3 subtype) was produced so that the H protein may secrete it to culture medium from the infected insect cells. The second viruses produced recombinant baculovirus which introduced cutting arrangement of factor-Xa into cell membrane attachment protein of the H protein. The protein was refined by affinity chromatography, ion exchangechromatography and gel filtration chromatography using specific antibody in order to large recover these 2 kinds of H proteins. With the goof the stage of the purification, the reactivity with the monoclonal antibody for measles virus lowered through the H protein. That H pro … More tein which reacts with the monoclonal antibody which shows only neutralizing antibody existed only for the fractionation in which electrophoresis and gel filtration chromatography recognize the large aggregation clarified. And, the activity which also hemagglutination of the green monkey red blood cells in either protein could not be recognized. The isolated purification of the protein from the recombinant baculovirus for structure analysis seems to be unsuitable from the above result. Therefore, it is concluded that the isolated purification of the protein from the virion is important in order to carry out three-dimensional structure analysis of the virus.And, we had already sensitized the measles virus in the gelatin particle, and the new measles antibody measuring methodwas developed. This was applied this time, and the measles IgM antibody measuring method was established. It was adopted for the measles eradication plan of WHO, because this method is convenient, and it would be examined in developing countries in Africa and South America, etc. Less