兔抗人多发性骨髓瘤细胞多克隆抗体的制备与鉴定

摘要

To prepare polyclonal antibody of rabbit anti-human multiple myeloma. Method:The New Zealand white rabbits were immunized with human multiple myeloma ARH-77 cell line at the back skin intradermal injection for the first time, he primary inoculation was with Freund's complete adju- vant (Sigma) ,follow by three boosts with Freund's incomplete adjuvant (Sigma) on the 14,28 and 38day. The serum was extracted after 4 ℃ settlement overnight. Polyclonal antibodies were isolated using affinity chromatography system. Purified polyclonal antibodies was analyzed by ELISA, Westem blot and Immunofluorescence stain. Results:polyclonal antibody titer was up to 1 : 20 000 by ELISA. Western blot showed that the antibody had a high specificity. Immunofluorescence stain showed Poly- clonal antibodies could specificitily binding with ARH-77 cell surface proteins. Conclusion:We pro- duced polyclonal antibodies of rabbit anti-human multiple myeloma and it have high titer and speci- ficity, to providing a new method for diagnosis and therapy of multiple myeloma.%制备人多发性骨髓瘤全细胞兔多克隆抗体。用人多发性骨髓瘤细胞系ARH-77全细胞和福氏完全佐剂通过背部皮内多点注射首次免疫新西兰大白兔,第14 d用ARH-77全细胞和福氏不完全佐剂同样剂量加强免疫,第28 d和38 d时再次免疫。第45 d颈动脉采血80 mL,4℃静置过夜,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。用酶联免疫吸附试验（ELISA）检测纯化多克隆抗体的效价;免疫印迹法和荧光免疫细胞化学检测纯化多克隆抗体的特异性。结果：ELISA检测多克隆抗体滴度为1∶20 000,免疫印迹检测结果显示该抗体具有较高特异性,荧光免疫细胞化学检测多克隆抗体能够有效的结合细胞表面抗原。实验获得了高效价的特异性的兔抗人骨髓瘤细胞多克隆抗体,为进一步研究多发性骨髓瘤诊断和治疗奠定了基础。