重组人α干扰素单克隆抗体的研制及其应用

Preparation and application of monoclonal antibodies to recombinant human IFN-α

目的 研制抗干扰素的单克隆抗体 (McAb)及其应用。方法 应用杂交瘤技术 ,获得 4 0株抗重组人α干扰素的单克隆抗体细胞株 ;用ELISA方法检测腹水滴度 ,用辛酸法纯化McAb。结果 4 0株细胞中 2E9、4G1能稳定分泌抗重组人α1b干扰素的单克隆抗体 ,2C9为抗重组新型干扰素 ,2A7、4G10分泌抗重组人α干扰素 (α1b、α2b、α2a、)和重组集成干扰素、重组新型干扰素的单克隆抗体细胞系 ;体外连续传代 6个月 ,分泌抗体能力不变 ,特异性专一。 5株单克隆抗体均为IgG。采用辛酸方法提纯抗体纯度达到 95 %以上。粗制的重组人α干扰素经 4G10单克隆抗体亲和层析柱提纯后 ,获得的干扰素纯度 95 %以上 ,平均收率 93% ,残余鼠IgG含量 <10 0ng 剂量 (5 0 μg)。 2C9单克隆抗体亲和层析柱适用于纯化新型干扰素。结论 4G10单克隆抗体亲和层析柱可以应用于大规模重组人α干扰素生产。

Objective To prepare and apply monoclonal antibodies (McAb) against recombinant human interferonalpha (rHu IFN α).Methods Five cell lines (2E9,4G1,2A7,2C9,4G10) secreting McAbs against rHu IFN α were established by hybridoma technique. Results All the cell lines secreted monoclonal antibodies stablely. Functions of secreting antibodies of the five cell lines lasted for 6 months in BALB/c mice and 8 months in cell culture. The specificity of antibody was constant. The Ig subclasses of the McAbs were IgG1. Anti IFN McAb affinity purification column was prepared by coupling the anti IFN α McAb to Sepharose 4B.The combining rate reached was higher than 95%. Conclusion The highest purification efficiency was obtained by using 4G10 column.