重组人白细胞介素10单克隆抗体的制备

Preparation of the monoclonal antibodies against recombinant human interleukin 10

目的 制备重组人白细胞介素 10 (rhIL 10 )单克隆抗体。方法 应用鼠 鼠杂交瘤技术建立分泌单克隆抗体的杂交瘤细胞系 ,ELISA检测杂交瘤细胞培养上清、腹水效价 ,双向免疫扩散试验进行亚类鉴定 ,Giemsa染色镜下计数杂交瘤染色体数目。结果 获得了 2株能分泌抗rhIL 10单克隆抗体的杂交瘤细胞系 (1D3、2A3) ,无血清培养液效价为 1∶5 12、1∶2 5 6 ,腹水效价 1∶10 5、1∶10 4 。单克隆抗体为IgG2 亚类 ,染色体数目 98～ 10 3条。 2株杂交瘤细胞体外传代培养6个月以上 ,ELISA检测抗体滴度无明显降低。结论 制备的 2株单克隆抗体效价高 。

Objective To prepare monoclonal antibodies (McAb) against recombinant human interleukin 10 (rhIL 10). Methods Mouse mouse hybridoma technolgy was used to set up hybridoma cell lines seereting McAbs. ELISA was used to detect the titer of serum free supernatant and ascetic fluid, double immunodiffusion test to analyse the immsunoglobulin subclass, et al . Results Two hybridoma cell lines (1D3, 2A3)secreting mcAbs were prepared, the titers of serum free supernatant were 1∶256～1∶512 and ascetic fluid were 1∶10 4～1∶10 5. The McAbs were IgG 2 subclass. Chromosomal numbers 98～103. The hybridoma cells were bequeathed more than six months and the titers of antibody did not change obviously. Conclusion The two McAbs against rhIL 10 show high titer and good reliability.