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目的：制备不同类型抗重组人血管生长素（Ｒｅｃｏｍｂｉｎａｎｔｉｏｎ Ｈｕｍａｎ Ａｎｇｉｏｇｅｎｉｎ ，ｒｈＡＮＧ） 的单克隆抗体，为进一步研究抗ｒｈＡＮＧ 抗体对ＡＮＧ 的中和作用以及对ＡＮＧ 依赖型肿瘤的抑瘤实验打下基础。方法：利用杂交瘤技术以ｒｈＡＮＧ 免疫Ｂａｌｂｃ小鼠，制备二株（Ｃ６ 、Ｃ４６） 能够稳定分泌抗ｒｈＡＮＧ 抗体的杂交瘤，并对上清进行ＥＬＩＳＡ、双扩散、免疫印迹等鉴定。结果：该抗体类别Ｃ６ 为ＩｇＧ１ ，Ｃ４６ 为ＩｇＭ，并特异地识别ｒｈＡＮＧ 与其它细胞因子及血清蛋白无交叉反应，免疫印迹显示该抗体与分子量１４４ｋＤｒｈＡＮＧ 蛋白结合，不与宿主菌体蛋白反应。结论：经多种方法鉴定获得的二株杂交瘤细胞分泌的抗体为ｒｈＡＮＧ 特异抗体。
OBJECTIVE: To prepare monoclonal antibodies against different types of recombinant human angiogenin (rhANG), and lay a foundation for the further study on the neutralizing effect of anti-rhANG antibody on ANG and the anti-tumor activity of ANG-dependent tumors. METHODS: Two hybridomas (C6 and C46) that could stably secrete anti-rhANG antibody were prepared by immunization of Balbc mice with hybridoma technology. The supernatants were identified by ELISA, double diffusion and Western blotting. Results: The antibody of C6 was IgG1, C46 was IgM, and specifically identified rhANG no cross-reaction with other cytokines and serum proteins. Western blotting showed that this antibody bound to the 14.4kD rhANG protein and did not react with host cell proteins. Conclusion: The antibodies secreted by the two hybridoma cells identified by various methods are rhANG-specific antibodies.