抗口蹄疫病毒非结构蛋白 3AB 的单克隆抗体的制备。
Generation of monoclonal antibodies against non-structural protein 3AB of foot-and-mouth disease virus.
机构信息
State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Vetersinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
出版信息
Virol Sin. 2012 Oct;27(5):316-9. doi: 10.1007/s12250-012-3261-x. Epub 2012 Oct 11.
To identify linear epitopes on the non-structural protein 3AB of foot-and-mouth disease virus (FMDV), BABL/c mice were immunized with the 3AB protein and splenocytes of BALB/c mice were fused with myeloma Sp2/0 cells. Two hybridoma monoclonal antibodies (mAbs) cell lines against the 3AB protein of foot-and-mouth disease virus (FMDV) were obtained, named C6 and E7 respectively. The microneutralization titer was 1:1024 for mAb C6, and 1:512 for E7. Both mAbs contain kappa light chains, and were of subclass IgG2b. In order to define the mAbs binding epitopes, the reactivity of these mAbs against FMDV were examined by indirect ELISA. The results showed that both mAbs can react with FMDV, but had no cross-reactivity with Swine Vesicular Disease (SVD) antigens. The titers in abdomen liquor were 1:5×10(6) for C6 and 1:2×10(6) for E7. In conclusion, the mAbs obtained from this study are specific for the detection of FMDV, can be used for etiological and immunological researches on FMDV, and have potential use in diagnosis and future vaccine designs.
为了鉴定口蹄疫病毒（FMDV）非结构蛋白 3AB 上的线性表位，用 3AB 蛋白免疫 BABL/c 小鼠，并用 BALB/c 小鼠的脾细胞融合骨髓瘤 Sp2/0 细胞。获得了针对口蹄疫病毒（FMDV）3AB 蛋白的两种杂交瘤单克隆抗体（mAb）细胞系，分别命名为 C6 和 E7。mAb C6 的微量中和效价为 1:1024，E7 为 1:512。两种 mAb 均含有κ轻链，亚类为 IgG2b。为了确定 mAb 的结合表位，通过间接 ELISA 检查了这些 mAb 对 FMDV 的反应性。结果表明，两种 mAb 均可与 FMDV 反应，但与猪水疱病（SVD）抗原无交叉反应性。C6 的腹水中滴度为 1:5×10(6)，E7 的腹水中滴度为 1:2×10(6)。综上所述，本研究获得的 mAb 特异性地检测到 FMDV，可用于 FMDV 的病因学和免疫学研究，具有在诊断和未来疫苗设计中的潜在用途。
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