Bruning J H, Debie W H, Dormans P H, Meijer H, Bruggeman C A
Arch Virol. 1987;94(1-2):55-70. doi: 10.1007/BF01313725.
In this paper the development of a battery of approximately 70 mouse monoclonal antibodies (McAbs) to RCMV-induced antigens and their characterization is discussed. Their reactivity with the whole scala of ca. 30 virus specific proteins was tested in an enzyme linked immunoassay (ELISA) whereas their ability to detect RCMV-antigens at different locations of in vitro infected cell cultures and at different stages of infection was tested by immunofluorescence. In order to determine to what specific (viral) protein each of these McAbs is directed against we used an immunoprecipitation technique, followed by SDS-PAGE. Furthermore, neutralizing capacity of each McAb was tested, as well as the immunoglobulin class they belong to. In this manner we defined six categories of monoclonal antibodies on the basis of immunofluorescence aspect. The six categories identify most important viral structural proteins.
本文讨论了一组约70种针对大鼠巨细胞病毒(RCMV)诱导抗原的小鼠单克隆抗体(McAbs)的研发及其特性。在酶联免疫吸附测定(ELISA)中测试了它们与约30种病毒特异性蛋白全谱的反应性,而通过免疫荧光测试了它们在体外感染细胞培养物的不同位置和不同感染阶段检测RCMV抗原的能力。为了确定这些单克隆抗体各自针对的是何种特异性(病毒)蛋白,我们采用了免疫沉淀技术,随后进行SDS-PAGE。此外,还测试了每种单克隆抗体的中和能力及其所属的免疫球蛋白类别。通过这种方式,我们根据免疫荧光特征定义了六类单克隆抗体。这六类抗体识别出了最重要的病毒结构蛋白。