鸡白细胞介素2分子单克隆抗体的研制

Development of monoclonal antibodies against chicken IL-2

以含有鸡白细胞介素2(ChIL-2)基因的重组真核表达质粒pcDNA3.1-ChIL-2免疫6周龄BALB/c小鼠,末次加强免疫后取其脾细胞与骨髓瘤细胞Sp2/0-Ag-14融合,应用间接ELISA筛选阳性克隆。共获得2株特异性分泌抗鸡IL-2分子的杂交瘤细胞株,分别命名为1H10和1E6,其腹水ELISA效价分别为1∶6400和1∶3200,亚类鉴定结果均为IgM。重组质粒pcDNA3.1-ChIL-2转染COS-7细胞,以上述单克隆抗体(mAb)检测表达产物,结果表明,2株mAb均能与表达产物反应;Western blot分析结果显示,2株mAb均与原核表达鸡IL-2蛋白发生特异性反应。本研究所制备的抗鸡IL-2分子mAb可为建立简单、快速的鸡IL-2检测方法提供可能,并为鸡IL-2生物学特性和禽类细胞免疫机理的研究提供材料。

Sp2/0-Ag-14 myeloma cells was fused with splenocytes from BALB/c mice immunized against chicken IL-2 and the supernatant of hybridoma clones were screened by indirect ELISA. Two hybridoma cell lines 1H10 and 1E6 secreting moloc-lonal antibodies （Mab） against ChlL-2 were obtained. The mAbs had an ascites titers pf 1：6 400 and 1：3 200 respectively and both were subtype IgM. IFA assay revealed that both mAbs could react with COS-7 cells transfected by pcDNA3.1-ChlL-2. Western blot analysis confirmed that these mAbs specifically recognized ChlL-2 only. Therefore the McAb developed in this work could be a useful tool to study avian immune cell function and immune regulations, and used as an effective reagent for the detection of chicken IL-2.