Brüning A, Phipps P, Posnett E, Canning E U
Department of Pure and Applied Biology, Imperial College of Science, Technology, and Medicine, London, UK.
Vet Parasitol. 1997 Jan;68(1-2):11-26. doi: 10.1016/s0304-4017(96)01074-6.
The production of monoclonal antibodies to the bloodstages of the haemoprotozoan parasites Babesia caballi and Babesia equi and the characterization of their corresponding antigens are described. Species specific and immunogenic proteins of both parasites were identified using SDS-PAGE, Western blotting and ELISA. These proteins were then electroeluted from SDS-PAGE gels and used to immunize BALB/c mice for hybridoma production. One monoclonal antibody (Mab), designated BC5.37.70.27 (BC5), recognized a 70 kDa protein of B. caballi as demonstrated by Western blotting under reducing conditions. Another Mab, BE1.24/2.95 (BEI), recognized a 34 kDa protein of B. equi. Both Mabs reacted specifically in indirect ELISA when isolated whole merozoites were used as antigen. Preliminary studies using the two Mabs in a competitive ELISA (cELISA) suggest that the cELISA for the detection of B. caballi infection is more sensitive than the commonly used complement fixation test but that refinement is necessary for the B. equi system.
本文描述了针对血液原虫寄生虫马巴贝斯虫（Babesia caballi）和马驽巴贝斯虫（Babesia equi）血液阶段的单克隆抗体的制备及其相应抗原的特性鉴定。使用SDS-PAGE、蛋白质印迹法和酶联免疫吸附测定（ELISA）鉴定了两种寄生虫的种特异性和免疫原性蛋白。然后从SDS-PAGE凝胶中电洗脱这些蛋白质，并用于免疫BALB/c小鼠以产生杂交瘤。一种单克隆抗体（Mab），命名为BC5.37.70.27（BC5），在还原条件下通过蛋白质印迹法证明可识别马巴贝斯虫的一种70 kDa蛋白。另一种Mab，BE1.24/2.95（BEI），可识别马驽巴贝斯虫的一种34 kDa蛋白。当使用分离的完整裂殖子作为抗原时，两种Mab在间接ELISA中均有特异性反应。使用这两种Mab进行竞争性ELISA（cELISA）的初步研究表明，用于检测马巴贝斯虫感染的cELISA比常用的补体结合试验更敏感，但马驽巴贝斯虫检测系统还需要进一步优化。
