麦长管蚜单克隆抗体及其特性研究

 【目的】制备麦长管蚜单克隆抗体，为进一步研究麦蚜的捕食性天敌种类奠定基础。【方法】应用杂交瘤技术制备单克隆抗体；双抗夹心酶联免疫吸附试验法（ELISA）鉴定抗体类型及亚类；间接ELISA测定抗体特异性；SDS-聚丙烯酰胺凝胶电泳和Western blot印迹分析确定抗体结合蛋白。【结果】制备出2株高度特异性的麦长管蚜单克隆抗体，分别命名为EGA-1E8和EGA-4H11，分别属于IgM亚类和IgG2b亚类。麦长管蚜抗原由多种多肽组成，其中单克隆抗体EGA-4H11与麦长管蚜抗原54.2 kD的多肽结合。特异性试验表明，这2种抗体不与麦田中其他种类的昆虫和蜘蛛发生交叉反应。【结论】成功地制备了麦长管蚜单克隆抗体，为进一步研究麦蚜的捕食性天敌种类奠定了可靠的基础。

The Monoclonal Antibodies to the English Wheat Aphid, Sitobion avenae (Fabricius), and Their Characteristics

【Objective】The English grain aphid, Sitobion avenae (Fabricius), is one of main wheat pests all over the world. It is very important to fully exert the natural control of predatory natural enemies in wheat fields in order to control the wheat aphid. 【Method】Hybridoma technolgy was used to develope monoclonal antibodies(McAbs) to the English grain aphid. The classes and subclasses of McAbs were determined by a double-antibody sandwich ELISA. Antibody specificity was tested using an indirect ELISA. SDS-PAGE and Western blot analysis were used to determine the proteins reacted with the antibodies.【Results】Two highly specific monoclonal antibodies were developed, namely EGA-1E8 and EGA-4H11, respectively. McAb EGA-1E8 was IgM subclass while McAb EGA-4H11 belonged to IgG2b subclass. McAb EGA-4H11 reacted with a single peptide of molecular weight 54.2 kD. The specificity test showed that both McAbs had no cross-reaction with other insects and spiders in wheat fields. 【Conclusion】The successful development of S. avenae McAbs may establish an essential tool for further study on the predatory natural enemies of the wheat aphid in wheat fields.