堆型艾美球虫子孢子单克隆抗体的制备及鉴定

Preparation and Identification of Monoclonal Antibody Against Sporozoites of Eimeria acervulina

目的建立堆型艾美球虫(Eimeriaacervulina)子孢子表面抗原的杂交瘤细胞株。方法使用纯化的堆型艾美球虫子孢子直接免疫BALB/c小鼠,采用杂交瘤技术建立细胞株,制备单克隆抗体。用ELISA确定单克隆抗体的交叉反应性、相对亲和力、免疫球蛋白的类型和亚类,并对单克隆抗体进行鉴定。结果获得4株可稳定分泌单克隆抗体的杂交瘤细胞株,其中Easp-3G3、Easp-5G10为IgG1类,Easp-3H6为IgG2b类,Easp-5H4为IgG2a类。4株单克隆抗体均能与堆型艾美球虫子孢子的抗原蛋白反应,但仅Easp-5H4能与柔嫩艾美球虫子孢子抗原蛋白反应。Easp-3G3和Easp-5G10与另外2种单克隆抗体的识别位点不同,而Easp-3G3与Easp-5G10、Easp-3H6与Easp-5H4的识别位点相近。结论制备了4株堆型艾美球虫子孢子单克隆抗体。

Objective To establish hybridoma cell lines against sperozoites of Eimeria acervulina. Methods BALB/c mice were immunized with purified sporozoites of E. acervulino. Hybridoma cell lines were set up by using hybridoma technique, and monoclonal antibodies were prepared. The monoclonal antibody was identified by determining their cross reactivity, relative affinity, immunoglobulin class or subclass with enzyme linked immunosorbent assay （ELISA）. Results Four hybridoma cell lines stably secreting McAbs against sporozoites were obtained： Easp-3G3 and Easp-5G10 belonging to IgG1, Easp-3H6 belonging to IgG2b, Easp-5H4 belonging to IgG2a. All four McAbs bound with E acervulina sporozoite protein, but the Easp-SH4 McAb showed cross reactivity withE teneUum sperozoite protein. Different antigenic epitopes were recognized by Easp-3G3 or Easp-5G10 and Easp-3H6 or Easp-5H4. Conclusion Three of the four produced monoclonal antibodies show high specificity and affinity to the Eimeria acervilina sporozoites.