Mäkelä M J, Norrby E, Salmi A
J Virol Methods. 1987 May;16(1-2):65-74. doi: 10.1016/0166-0934(87)90032-2.
A competitive enzyme immunoassay (EIA) for measuring polypeptide- and antigenic site-specific antibodies against measles virus (MV) was developed. Three monoclonal antibodies (MAbs) against the fusion (F) and one MAb against the hemagglutinin (H) protein of MV were labelled with biotin and used in biotin-streptavidin EIA system. The inhibition of the binding of the labelled MAbs by the bound serum samples was measured in EIA and the titers were defined by reading at the level of 25% inhibition. The titers increased from acute to early convalescent measles sera in all the tests and antibody levels to the F protein increased earlier than antibodies to the H protein. The ratio of antibodies against 2 different antigenic sites on the F protein was significantly higher in subacute sclerosing panencephalitis patients than in normal blood donors. Similarly, SJL mice immunized with MV antigen had higher ratios of the corresponding antibodies than Balb/c mice. The results indicate that antibody response to different polypeptides and antigenic sites on the same polypeptide can be studied with the developed method and that the responses against different antigenic sites are independent.
开发了一种用于检测针对麻疹病毒(MV)的多肽和抗原位点特异性抗体的竞争性酶免疫测定(EIA)。三种针对MV融合蛋白(F)的单克隆抗体(MAb)和一种针对MV血凝素(H)蛋白的MAb用生物素标记,并用于生物素-链霉亲和素EIA系统。在EIA中测量结合的血清样品对标记MAb结合的抑制作用,并通过在25%抑制水平下读数来确定滴度。在所有测试中,滴度从急性麻疹血清到早期恢复期麻疹血清均升高,并且针对F蛋白的抗体水平比针对H蛋白的抗体升高得更早。亚急性硬化性全脑炎患者针对F蛋白上2个不同抗原位点的抗体比率显著高于正常献血者。同样,用MV抗原免疫的SJL小鼠比Balb/c小鼠具有更高的相应抗体比率。结果表明,利用所开发的方法可以研究对不同多肽以及同一多肽上不同抗原位点的抗体反应,并且针对不同抗原位点的反应是独立的。