Balu S, Rothwell L, Kaiser P
Institute for Animal Health, Compton, Berkshire RG20 7NN, UK.
Vet Immunol Immunopathol. 2011 Mar 15;140(1-2):140-6. doi: 10.1016/j.vetimm.2010.11.009. Epub 2010 Nov 18.
Using genetic immunisation of mice, we produced antibodies against chicken interleukin-12p40 (chIL-12p40), also known as IL-12β. After a final injection with a recombinant chIL-12p40 protein, several stable hybridoma cell lines were established which secreted monoclonal antibodies (mAbs) to this component of the heterodimeric IL-12 cytokine. Specific binding of three of the mAbs to COS-7 cell-derived recombinant chIL-12p40 and the chIL-12p70 heterodimer was demonstrated in an indirect ELISA, and in dot blots. Two of the mAbs were used to develop a capture ELISA, suitable for detecting both recombinant protein (chIL-12p40 and the heterodimeric p70 protein) and native chIL-12. The mAbs were further characterised to show utility in immunocytochemistry.
通过对小鼠进行基因免疫，我们制备了针对鸡白细胞介素-12p40（chIL-12p40，也称为IL-12β）的抗体。在用重组chIL-12p40蛋白进行最后一次注射后，建立了几个稳定的杂交瘤细胞系，这些细胞系分泌针对异二聚体白细胞介素-12细胞因子这一成分的单克隆抗体（mAb）。在间接ELISA和斑点印迹中证实了三种mAb与COS-7细胞衍生的重组chIL-12p40和chIL-12p70异二聚体的特异性结合。其中两种mAb被用于开发一种捕获ELISA，适用于检测重组蛋白（chIL-12p40和异二聚体p70蛋白）和天然chIL-12。对这些mAb进行了进一步表征，以显示其在免疫细胞化学中的实用性。
