抗人白细胞间素-2多克隆及单克隆抗体的生物学特征

利用人工合成的白细胞间素2(IL-2)多肽片段,用中和试验、竞争ELISA和染色IL-2分泌细胞的碱性磷酸酶抗碱性磷酸酶桥联酶标技术,对己制备的抗人IL-2多克隆抗体及单克隆抗体(McAb)的表位特异性、中和活性、亲和常数及杭天然IL-2的反应性进行了全面鉴定。结果表明:利用多种免疫方案、融合前强化加强免疫及多个脾脏混合融合制备策略所获得的McAb,在生物学特征上表现出多样化,即这些McAb的表位特异性、中和活性、亲和力及抗天然IL-2活性各不相同。其中抗IL-2 C末端的McAb为首次报道。这组McAb适用于对IL-2研究的不同目的。

Biological Characteristics of Polyclonal and Monoclonal-Antibodies against Human Interleukin-2

Polyclonal-and monoclonal-antibodies ( PcAb and McAb ) against human interleukin-2 were characterized by their epitope specificities, affinities and reactivities to recombinant and natural interleukin-2 by using 1) synthesized IL-2 peptides; 2) a competitive ELISA and 3) an IL-2-containing cell staining APAAP method. The results showed that, by our used strategies in making hybridomas ( varied immunizing methods, enhanced boostings and using mixed spleen cells for fusion ) , monoclonal antibodies with various biological characteristics have been obtained. Among these McAbs, four were found to have specificities to the N-terminals of IL-2 molecules, 4) to the 59-72 residuses, 2 to the C-terminals of IL-2 molecules and 2 with uncertain epitope specificities. The McAbs with neutralizing activities were mainly those having the epitope-specificities to the N-terminals of the IL-2 molecules. The McAbs reacted with natural IL-2 were also identified. These antibodies could be designed io have different uses.