兔抗小鼠多发性骨髓瘤细胞多克隆抗体的制备及抗肿瘤研究

目的:制备小鼠多发性骨髓瘤全细胞兔多克隆抗体,并初步研究其抗肿瘤活性。方法:用小鼠多发性骨髓瘤细胞MPC-11活细胞通过背部皮内多点注射多次免疫新西兰大白兔,收集血清。然后采用亲和层析系统纯化血清中的多克隆抗体。用ELISA检测纯化多克隆抗体的效价,MTT法分析多克隆抗体对MPC-11增殖的影响,通过皮下肿瘤模型研究多克隆抗体对荷瘤小鼠体内肿瘤生长的影响。结果:我们获得了高特异性的多克隆抗体,ELISA检测抗体滴度在1∶20 000以上,MTT研究发现200μg/mL多克隆抗体处理骨髓瘤细胞MPC-11 48 h后,明显抑制骨髓瘤细胞生长;体内研究发现,多克隆抗体明显抑制小鼠皮下肿瘤的生长。结论:实验获得了高效价的特异性兔抗小鼠骨髓瘤细胞多克隆抗体,通过体内体外研究发现,多克隆抗体能够明显抑制肿瘤细胞的生长,为进一步研究多发性骨髓瘤的诊断和治疗提供了新的方法。

Preparation and anti-tumor research of polyclonal antibodies of rabbit anti-mouse multiple myeloma cells

Objective:To prepare rabbit polyclonal antibodies with mouse multiple myeloma cells and study its antitumor activity.Methods: The polyclonal antibody was generated by multi-point immunizing New Zealand white rabbits with live MPC-11 cells.The serum was pooled a week after the last injection.Immunoglobulin was isolated for using an affinity chromatography system and then freeze dried.Elisa was employed to detect and identify the specific binding of PAb and inhibited multiple myeloma cell proliferation as revealed by MTT assay.Serial intravenous injections of PAb inhibited tumor growth in mice bearing murine multiple myeloma cells.Results: Highly specific polyclonal antibodwere obtaines.ELISA detection found that antibody titers were more than 1∶ 20 000,The MTT study found that the cytotoxicity of PAb on MPC-11 cell lines was both dose-dependent and time-dependent.The PAb exerted a 50% inhibitory effect on MPC-11 cell viability at a concentration of 200.0 μg/mL in 48 h and the polyclonal antibody significantly inhibited the growth of subcutaneous tumors in mice by in vivo study.Conclusion: High specific rabbit anti-mouse myeloma cell polyclonal antibody can be obtrained.The study results show that the polyclonal antibody can significantly inhibit tumor cell growth in vitro and in vivo,The data also suggests that PAb is an effective agent for multiple myeloma therapy and that exploratory clinical trials may be warranted.