Stratieva-Taneeva P A, Khaidukov S V, Kovalenko V A, Nazimov I V, Samokhvalova L V, Nesmeyanov V A
Shemyakin Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
Hybridoma. 1993 Jun;12(3):271-84. doi: 10.1089/hyb.1993.12.271.
Hybrid hybridomas (tetradomas), producing bispecific monoclonal antibodies (bmabs) binding to both interleukin 2 and horseradish peroxidase were obtained by fusing IL 2-specific and HRP-specific hybridomas. Parental hybridomas were labelled prior to the fusion with fluorescein isothiocyanate and tetramethylrhodamine isothiocyanate. Bifluorescent (tetradoma) cells were sorted out using fluorescence activated cell sorter. Two clones, designated D8C1/G and H7C11/H were shown to secrete bmabs over at least 6 months growth in culture. Bmabs have been purified from mouse ascites by ammonium sulfate precipitation and affinity chromatography on immobilized peptide, modelling corresponding IL 2 epitope. The purity of antibodies obtained was characterized by capillary electrophoresis. The possibility of using these antibody preparations in rIL 2 analysis was evaluated in two types of EIA: direct and competitive solid-phase EIA. Both assays had similar sensitivity of about 1.5-4 ng IL 2 per ml.
通过将白细胞介素2特异性杂交瘤和辣根过氧化物酶特异性杂交瘤进行融合，获得了能产生与白细胞介素2和辣根过氧化物酶均结合的双特异性单克隆抗体（双特异性抗体）的杂交杂交瘤（四联瘤）。亲代杂交瘤在融合前用异硫氰酸荧光素和异硫氰酸四甲基罗丹明进行标记。使用荧光激活细胞分选仪筛选出双荧光（四联瘤）细胞。在培养至少6个月的过程中，两个命名为D8C1/G和H7C11/H的克隆显示出能分泌双特异性抗体。双特异性抗体已通过硫酸铵沉淀和固定化肽亲和色谱法从小鼠腹水中纯化出来，该肽模拟相应的白细胞介素2表位。通过毛细管电泳对所获得抗体的纯度进行了表征。在两种酶免疫分析（EIA）类型中评估了使用这些抗体制剂进行重组白细胞介素2分析的可能性：直接和竞争性固相EIA。两种检测方法的灵敏度相似，约为每毫升1.5 - 4纳克白细胞介素2。
