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目的利用纯化的重组人肝再生 增强因子(hALR)制备抗hALR的单克隆抗体,建立hALR的检测方法。方法采用杂交瘤技术制备单克隆抗体,经ELISA和免疫印迹证明其特异性。 结果获得了4株分泌抗hALR特异性抗体的杂交瘤细胞 系;无血清培养液效价为1×10~(-2),腹水效价为1×10~(-5);亚类鉴定 表明:2株单克隆抗体为IgG1,另2株为IgG2b;免疫印迹显示:抗体结合抗原的分子量 与hALR相符,为特异性抗hALR抗体。结论通过杂交瘤 技术,获得了抗hALR的单克隆抗体,为以后的工作奠定了基础。
OBJECTIVE: To prepare monoclonal antibodies against hALR by using purified recombinant human augmenter of liver regeneration (hALR) and to establish a method for detecting hALR. Methods Monoclonal antibodies were prepared by hybridoma technique and their specificity was confirmed by ELISA and Western blotting. Results Four hybridoma cell lines secreting anti-hALR specific antibodies were obtained. The titer of serum-free medium was 1 × 10 -2 and the titer of ascites was 1 × 10 -5. : Two monoclonal antibodies were IgG1 and the other two were IgG2b; Western blotting showed that the molecular weight of the antibody binding antigen was consistent with hALR, which was a specific anti-hALR antibody. Conclusion The monoclonal antibody against hALR was obtained by hybridoma technology, which laid the foundation for future work.