人骨肉瘤OS-732单克隆抗体的制备及纯化

Preparation and purification of murine monoclonal antibody against human osteosarcoma OS-732

摘要: 目的制备、纯化抗人骨肉瘤OS-732单克隆抗体。方法通过杂交瘤细胞腹腔注射BALB/c小鼠制备腹水,ProteinA-Sepharose CL-4B亲和层析法纯化单抗,SDS-PAGE、免疫组化及ELISA检测该单抗的性质。结果 SDS-PAGE鉴定单抗纯度达到93%,ELISA检测抗体免疫活性效价为1×10^-7。结论制备、纯化了具有较高纯度及生物活性的的抗骨肉瘤OS-732单克隆抗体,为下一步骨肉瘤靶向治疗打下了基础。

Abstract: Objective To prepare and purify the murine antibody against human osteosarcoma OS-732. Methods Human osteosarcoma OS-732 and hybridoma cells were injected into the abdominal cavity of BALB/c mice to produce monoclonal antibody against human osteosarcoma.The antibody was purified by protein A-sepharose CL-4B affinity chromatography and its properties were evaluated by SDS-PAGE,immunohistochemistry and ELISA. Results SDS-PAGE showed that the purity of monoclonal antibody was about 93%.ELISA revealed that the immunologic activity of monoclonal antibody was 1×10-7. Conclusion Monoclonal antibodies with a high purity and immunological activity can be employed in further study of targeted therapy for human osteosarcoma.