重组人α2a型干扰素单克隆抗体的研制、特性鉴定及初步应用

Preparation,Characterization and application of monoclonal antibodies to recombinant human interferon—α2a

应用常规方法建立了5株稳定分泌抗重组人α2a 型干扰素单克隆抗体的小鼠杂交瘤细胞系1B8、2B6、3F1、4G5和5G10,并对其免疫学特性进行了较系统的鉴定.5株单抗均为 IgG1,特异性强,与重组人α1 型、γ型干扰素以及受体菌菌体蛋白成分无交叉反应.间接 ELISA 测定小鼠腹水单抗效价1:320,000～1:10,240,000,其中3F1和5G10单抗对重组人α2a 型干扰素具中和活性(中和效价均为1:10,000)。竞争结合 ELISA 证实5株单抗针对3种不同表位。用3F1和5G10单抗建立的夹心 ELISA 能检测到60pg/ml的重组人α2a 和α2b 型干扰素。3F1单抗用于制备亲和层析柱纯化重组人α2a 型干扰素结果与进口单抗NK2相当。

In this report,five mouse hybridoma cell lines secreting monoclonal antibodies to recombi-nant human interferon—α2a(rHu IFN—α2a)were raised.All the monoclonal antibodies with IgGl subclass react with rHu IFN—α2a detected by indirect ELISA with the titers ranging from 1:320,000 to 1:10,240,000 but have no cross reactivity with rHu IFNαl,γ and proteins from recombinant E.coli.Two of monoclonal antibodies,designated 3F1 and 5G10,have the ability to neutralize antiviral activity of rHu IFN—α2a.By a competitive ELISA the five monoclonal anti-bodies can be divided into three categories based on their epitope specificities.A sandwich ELISA established by employing 3F1 and 5G10 monoclonal antibodies could detect rHuIFN-α2a or rHuIFN-α2b as few as 60pg/ml·Moreover,3F1 monoclonal antibody displayed a satisfactory property in affinity chromatography for rHuIFNα2a,which is able to match a monoclonal anti- body NK2 from CellTech Corporation.