沙拉沙星单克隆抗体制备及鉴定  

Preparation and Identification of Monoclonal Antibody for Sarafloxacin

作　　者：刘儒彪[1] 胡骁飞[2] 张改平[2,3] 王慧[1] 王方雨[2] 刘璐[1] 刘玺[1] 邓瑞广[2] 宋照军[1] 

机构地区：[1]河南科技学院食品学院,河南新乡453003 [2]河南省农业科学院动物免疫学重点实验室,河南郑州450002 [3]河南农业大学牧医工程学院,河南郑州450002

出　　处：《华北农学报》2016年第2期87-91,共5页Acta Agriculturae Boreali-Sinica

基　　金：2015年河南省科技攻关项目(152102310094);河南省高校科技创新团队支持计划项目(13IRTSTHN006)

摘　　要：为制备敏感性好、亲和力高、特异性强的抗沙拉沙星（Sarafloxacin,SARA）单克隆抗体,采用碳二亚胺法合成SARA人工抗原,通过免疫BALB/c小鼠,选择血清效价高且敏感性好的小鼠,采用杂交瘤细胞技术进行细胞融合,筛选分泌抗SARA单克隆抗体（Monoclonal antibody,mAb）的杂交瘤细胞株;采用体内诱生腹水法制备SARA mAb,通过间接ELISA和间接竞争ELISA对SARA mAb的免疫学特性进行鉴定。结果表明：小鼠经免疫原免疫后,小鼠多抗血清效价均达到1∶128 000。选择敏感性较好的2号小鼠进行细胞融合,经多次亚克隆后,筛选出1G3、3H3两株杂交瘤细胞株,其中1G3所产SARA mAb效价较高,为1∶512 000,IC_（50）为6.34 ng/mL,亲和常数为8.55×10~8L/mol,与诺氟沙星的交叉反应率为1.06%,与其他药物交叉反应率均低于0.50%。In order to preparation of sarafloxacin（ SARA） monoclonal antibody with good sensitivity,high affinity and strong specificity,synthesized SARA artificial antigen by carbodiimide method. It was used to immunize BALB / c mice. The mice which possessed high serum titer and good sensitivity were selected for cell fusing through cell hybridoma technique. Then to filtrate the hybridoma cell strain which secreted SARA monoclonal antibody（ mAb）. Prepared SARA mAb by inducing ascites in vivo,and identificated the immunity characteristics of SARA mAb via indirect ELISA and indirect competitive ELISA. The result indicated that after mice were immune by immunogen,their serum antibody titer were all up to 1 ∶ 128 000. Number 2 mouse with good sensitivity was chosen for cell fusion. After multiple sub cloning,1G3 and 3H3 hybridoma cell strain were filtered. The titer of SARA mAb from 1G3 was 1 ∶ 512 000,IC_（50）was 6. 34 ng / m L,affinity constant was 8. 55 × 108 L / mol,cross-reactivity with norfloxacin was 1. 06%,cross-reactivity with other drugs were below 0. 50%,higher than that of 3H3.

关 键 词：沙拉沙星 单克隆抗体 杂交瘤细胞 免疫学特性 

分 类 号：S859.84[农业科学—临床兽医学]