抗成骨肉瘤mAb的制备、鉴定及其细胞毒效应

来源 :细胞与分子免疫学杂志 | 被引量 : 0次 | 上传用户:bingdongfenxing
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目的建立抗人成骨肉瘤mAb杂交瘤细胞系并对其抗体特性进行研究。方法利用我室自建的骨肉瘤细胞系(OS-9607)免疫Balb/c小鼠,取小鼠脾细胞与骨髓瘤细胞Sp2/0进行融合,经筛选和克隆化后,建立杂交瘤细胞系。以免疫组化和Western Blot等方法研究抗体特性,以MTT法研究细胞毒作用。结果得到1株能够稳定分泌抗体,效价高的杂交瘤细胞系3D9,免疫组化结果显示着色区主要分布在细胞核上,相应抗原在成骨肉瘤标本和成骨肉瘤细胞系高度表达(83%),正常组织未见表达,Western Blot 显示3D9相应分子量为Mr 54 000。MTT法示该抗体对成骨肉瘤细胞的杀伤率明显高于对照组。结论得到的杂交瘤细胞系分泌的mAb体具有成骨肉瘤细胞特异亲和性,并有一定的细胞毒效应,可能成为新的成骨肉瘤生化标志,在肿瘤的免疫治疗方面有广阔的应用前景。 Objective To establish an anti-human osteosarcoma mAb hybridoma cell line and study its antibody characteristics. Methods Balb / c mice were immunized with self-built osteosarcoma cell line OS-9607 and splenocytes of mouse were fused with Sp2 / 0 myeloma cells. After screening and cloning, the hybridoma cell lines . Immunohistochemistry and Western Blot and other methods to study the characteristics of antibodies, MTT assay cytotoxicity. Results A highly competent hybridoma cell line, 3D9, was obtained. The results of immunohistochemistry showed that the staining region was mainly located in the nucleus and the corresponding antigen was highly expressed in osteosarcoma specimens and osteogenic sarcoma cell lines (83% ), No expression in normal tissues, Western Blot showed that the corresponding molecular weight of 3D9 Mr 54 000. MTT method showed that the antibody killing rate of osteosarcoma cells was significantly higher than the control group. Conclusions The mAb secreted by the hybridoma cell line has the specific affinity of osteogenic sarcoma cells and has certain cytotoxic effect and may be a new biochemical marker of osteosarcoma. It has broad application prospects in the immunotherapy of tumors .
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