Wang Cheng-Min, He Hong-Xuan, Qin Jian-Hua, Chen Gui-Xiang, Guo Qi-Zhen, Hang Bo-Lin
Cell Engineering Key Laboratory of Henan Institute of Science and Technology, Xinxiang 453003, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2006 Dec;24(6):453-6.
To establish hybridoma cell lines against sporozoites of Eimeria acervulina.
BALB/c mice were immunized with purified sporozoites of E. acervulina. 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).
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-5H4 McAb showed cross reactivity with E. tenellum sporozoite protein. Different antigenic epitopes were recognized by Easp-3G3 or Easp-5G10 and Easp-3H6 or Easp-5H4.
Three of the four produced monoclonal antibodies show high specificity and affinity to the Eimeria acervulina sporozoites.
建立抗堆型艾美耳球虫子孢子的杂交瘤细胞系。
用纯化的堆型艾美耳球虫子孢子免疫BALB/c小鼠。采用杂交瘤技术建立杂交瘤细胞系,并制备单克隆抗体。通过酶联免疫吸附测定(ELISA)测定其交叉反应性、相对亲和力、免疫球蛋白类别或亚类来鉴定单克隆抗体。
获得了4株稳定分泌抗子孢子单克隆抗体的杂交瘤细胞系:Easp-3G3和Easp-5G10属于IgG1,Easp-3H6属于IgG2b,Easp-5H4属于IgG2a。这4种单克隆抗体均与堆型艾美耳球虫子孢子蛋白结合,但Easp-5H4单克隆抗体与柔嫩艾美耳球虫子孢子蛋白有交叉反应。Easp-3G3或Easp-5G10与Easp-3H6或Easp-5H4识别不同的抗原表位。
所产生的4种单克隆抗体中有3种对堆型艾美耳球虫子孢子具有高度特异性和亲和力。