山羊支原体山羊肺炎亚种单克隆抗体的制备及抗体结合抗原表位的筛选  

Preparation of Monoclonal Antibodies against Mycoplasma capricolum subsp.capripneumoniae and Screening of Antibody Binding Epitopes

作　　者：郑莹莹[1] 骈亚亚[2] 储岳峰[1] 赵萍[1] 贺英[1] 简莹娜 姜永强[2] 逯忠新[1] 

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部草食动物疫病重点开放实验室,农业部畜禽病毒学重点开放实验室,甘肃兰州730046 [2]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071

出　　处：《中国畜牧兽医》2014年第8期21-24,共4页China Animal Husbandry & Veterinary Medicine

基　　金：甘肃省重大科技专项

摘　　要：本研究旨在制备山羊支原体山羊肺炎亚种(M.capricolumsubsp.capripneumoniae,Mccp)的单克隆抗体并筛选与抗体结合的抗原表位。试验用甲醛灭活的Mccp免疫BALB/c小鼠,运用传统的细胞融合技术进行融合获得杂交瘤细胞,亚克隆,制备单克隆抗体腹水,采用酶联免疫技术(ELISA)和免疫印迹(Western blotting)技术鉴定单克隆抗体的特异性及胶内酶切鉴定与抗体结合的抗原表位。最终成功筛选获得5株单克隆抗体,鉴定到3个与抗体结合的抗原表位,为下一步科学研究提供了初步且可靠的试验基础。The aim of this research was to prepare monoclonal antibodies against Mycoplasma capricolum subsp.capripneumoniae（M.capricolumsubsp.capripneumoniae,Mccp）,and also screen antibody binding epitopes.We inactivated Mccp with formaldehyde,and then immunized BALB/c mice.We applied conventional fusion cell technology to obtain hybridoma cell,subclones,and ascites of monoclonal antibodies.We used enzyme-linked immunoassay（ELISA）and immunoblotting（Western blotting）technologies to identify the specificity of monoclonal antibodies,and enzyme digestion method to identify antibody binding epitopes.Finally,we successfully screened five monoclonal antibodies and identified three antibody binding epitopes,which provided a preliminary and reliable experimental basis for further scientific research.

关 键 词：山羊支原体山羊肺炎亚种 单克隆抗体 抗体结合的抗原表位 

分 类 号：Q78[生物学—分子生物学]