Emini E A, Jameson B A, Lewis A J, Larsen G R, Wimmer E
J Virol. 1982 Sep;43(3):997-1005. doi: 10.1128/JVI.43.3.997-1005.1982.
Two hybridomas (H3 and D3) secreting monoclonal neutralizing antibody to intact poliovirus type 1 (Mahoney strain) were established. Each antibody bound to a site qualitatively different from that to which the other antibody bound. The H3 site was located on intact virions and, to a lesser extent, on 80S naturally occurring empty capsids and 14S precursor subunits. The D3 site was found only on virions and empty capsids. Neither site was expressed on 80S heat-treated virions. The antibodies did not react with free denatured or undenatured viral structural proteins. Viral variants which were no longer capable of being neutralized by either one or the other antibody were obtained. Such variants arose during normal cell culture passage of wild-type virus and were present in the progeny viral population on the order of 10(-4) variant per wild-type virus PFU. Toluene-2,4-diisocyanate, a heterobifunctional covalent cross-linking reagent, was used to irreversibly bind the F(ab) fragments of the two antibodies to their respective binding sites. In this way, VP1 was identified as the structural protein containing both sites.
建立了两种分泌针对完整1型脊髓灰质炎病毒（马奥尼株）的单克隆中和抗体的杂交瘤（H3和D3）。每种抗体结合的位点在性质上与另一种抗体结合的位点不同。H3位点位于完整病毒粒子上，在较小程度上也位于80S天然存在的空衣壳和14S前体亚基上。D3位点仅在病毒粒子和空衣壳上发现。两个位点在80S热处理病毒粒子上均未表达。这些抗体不与游离的变性或未变性病毒结构蛋白反应。获得了不再能被其中任何一种抗体中和的病毒变体。这种变体在野生型病毒的正常细胞培养传代过程中出现，并且在子代病毒群体中的存在比例约为每野生型病毒PFU有10(-4)个变体。使用甲苯-2,4-二异氰酸酯，一种异双功能共价交联试剂，将两种抗体的F(ab)片段不可逆地结合到它们各自的结合位点。通过这种方式确定VP1为包含这两个位点的结构蛋白。
