Schreier E, Stompor S, Michel S
Department of Molecular Biology, Central Institute of Hygiene, Microbiology and Epidemiology, Berlin, G.D.R.
Acta Virol. 1988 Jul;32(4):296-302.
The polymerase proteins (PB1, PB2, PA) of the influenza virus strain A/PR/8/34 (H1N1) were isolated from whole virion or ribonucleoprotein (RNP) fractions by electrophoresis on polyacrylamide gel and electroelution or Sepharose CL-6B chromatography in the presence of SDS. Antisera to polymerase proteins (P proteins) were raised in rabbits; the immunoglobulins (Ig) were purified by affinity chromatography. Characterization of the antibody fraction by Western blot analysis showed a highly monospecific reaction with the three polymerase proteins. Spot immunobinding assay was used to compare the immunoreactivity of the monospecific polymerase antibodies with the P proteins of other influenza A subtypes and influenza B strains, revealing high immunoreactivity with the components of all influenza A strains and only insignificant reactivity with the components of the influenza B strains tested.
通过在十二烷基硫酸钠(SDS)存在的条件下,在聚丙烯酰胺凝胶上进行电泳以及电洗脱或用琼脂糖凝胶CL-6B进行色谱分析,从全病毒体或核糖核蛋白(RNP)组分中分离出甲型流感病毒A/PR/8/34(H1N1)株的聚合酶蛋白(PB1、PB2、PA)。用兔制备针对聚合酶蛋白(P蛋白)的抗血清;通过亲和色谱法纯化免疫球蛋白(Ig)。通过蛋白质印迹分析对抗体组分进行表征,结果显示其与三种聚合酶蛋白发生高度单特异性反应。采用斑点免疫结合试验比较单特异性聚合酶抗体与其他甲型流感病毒亚型及乙型流感病毒株P蛋白的免疫反应性,结果显示其与所有甲型流感病毒株的组分具有高免疫反应性,而与所检测的乙型流感病毒株的组分仅有微弱反应。