黄热病病毒糖蛋白的抗原分析：单克隆抗体在酶联免疫吸附测定中的应用

Antigenic analysis of yellow fever virus glycoproteins: use of monoclonal antibodies in enzyme-linked immunosorbent assays.

作者信息

Cammack N, Gould E A

出版信息

J Virol Methods. 1986 May;13(2):135-42. doi: 10.1016/0166-0934(86)90081-9.

Abstract

A sensitive enzyme-linked immunosorbent assay with biotin and streptavidin (B/SA ELISA) was developed for the specific detection of yellow fever (YF) viruses. Monoclonal antibodies (MCA) specific for the envelope (E) glycoprotein or the non-structural glycoprotein (NV3) of YF virus-infected cell lysates were tested by antigen and antibody capture B/SA ELISA against YF viruses and a large number of other flaviviruses. In an antigen capture assay, with suckling mouse brain antigens, MCA directed against the envelope glycoprotein clearly differentiated YF viruses from any other flaviviruses, wild-type YF viruses from YF vaccine viruses and YF17D-204 substrains from other YF vaccine viruses. Specific MCA could identify YF viruses in an antibody capture assay with concentrated YF-infected tissue culture supernatant medium as the solid phase. On the basis of the principles established above, MCA binding profiles of purified YF virus strains were compared. No qualitative differences in immunoreactivity could be detected between closely related strains of YF virus. An antibody capture assay on infected tissue culture monolayers was developed to enable in situ detection of YF viruses using MCA specific for both the envelope glycoprotein and the non-structural NV3 glycoprotein.

摘要

开发了一种采用生物素和链霉亲和素的灵敏酶联免疫吸附测定法（B/SA ELISA），用于特异性检测黄热病毒。针对黄热病毒感染细胞裂解物的包膜（E）糖蛋白或非结构糖蛋白（NV3）的单克隆抗体（MCA），通过抗原和抗体捕获B/SA ELISA，针对黄热病毒和大量其他黄病毒进行了检测。在抗原捕获试验中，使用乳鼠脑抗原，针对包膜糖蛋白的MCA能清楚地将黄热病毒与任何其他黄病毒区分开来，将野生型黄热病毒与黄热疫苗病毒区分开来，将YF17D - 204亚株与其他黄热疫苗病毒区分开来。特异性MCA可以在以浓缩的黄热病毒感染组织培养上清液为固相的抗体捕获试验中识别黄热病毒。基于上述确立的原理，比较了纯化的黄热病毒株的MCA结合谱。在密切相关的黄热病毒株之间未检测到免疫反应性的定性差异。开发了一种针对感染组织培养单层细胞的抗体捕获试验，以使用针对包膜糖蛋白和非结构NV3糖蛋白的MCA进行黄热病毒的原位检测。