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目的制备并纯化具有高特异性的抗星形胶质细胞上调基因1(AEG-1)单克隆抗体(mAb),并进行异硫氰酸荧光素(FITC)标记,鉴定标记的mAb与肿瘤细胞的结合能力。方法小鼠腹水诱生法制备并纯化抗AEG-1 mAb,Western blot法和免疫组织化学染色检测mAb的纯度及免疫活性。流式细胞术检测FITC标记mAb的标记效率,流式细胞术及免疫细胞化学染色检测FITC标记mAb与肿瘤细胞的结合能力。结果纯化得到了具有较高纯度的抗AEG-1 mAb,免疫活性良好;流式细胞术检测结果显示,FITC标记的mAb标记的肿瘤细胞表面荧光强度明显增加,标记率可达到90%以上;FITC标记的mAb可特异性识别并结合AEG-1阳性表达的肿瘤细胞,还能与小鼠体内注射的肿瘤细胞结合,在BALB/c小鼠血液中可检测出荧光染色阳性的肿瘤细胞。结论成功制备出小鼠抗AEG-1 mAb并进行了FITC标记,标记的mAb在体内外均可与肿瘤细胞特异性结合。
Objective To prepare and purify highly specific anti-astrocyte up-regulated gene 1 (mAb) monoclonal antibody (mAb) and fluorescein isothiocyanate (FITC) labeling, identification of labeled mAb and tumor cells The ability to combine. Methods Anti-AEG-1 mAb was prepared and purified by mouse ascites induction method. The purity and immunological activity of mAb were detected by Western blot and immunohistochemical staining. Flow cytometry was used to detect the labeling efficiency of FITC-labeled mAb. Flow cytometry and immunocytochemical staining were used to detect the binding ability of FITC-labeled mAbs to tumor cells. Results The anti-AEG-1 mAb with high purity was purified and the immunological activity was good. The results of flow cytometry showed that the fluorescence intensity of FITC-labeled mAb markedly increased and the labeling rate could reach more than 90%. FITC The labeled mAb specifically recognizes and binds to AEG-1-positive tumor cells and binds to tumor cells injected in vivo. Fluorescent stain-positive tumor cells can be detected in the blood of BALB / c mice. Conclusion The mouse anti-AEG-1 mAb was successfully prepared and labeled with FITC. The labeled mAb could specifically bind to tumor cells both in vitro and in vivo.