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当前位置: 首页 > 文章 > 环丙沙星单克隆抗体的制备及鉴定 食品科学 2010,31 (12) 16-20
Position: Home > Articles > Preparation and Identification of Monoclonal Antibody against Ciprofloxacin FOOD SCIENCE 2010,31 (12) 16-20

环丙沙星单克隆抗体的制备及鉴定

作  者：
杨先乐;黄宣运;胡鲲;方伟;王炼;姜有声
单  位：
上海海洋大学;四川省水产学校
关键词：
环丙沙星;半抗原;抗体;免疫
摘  要：
通过改进的碳二亚胺法制备环丙沙星-牛血清白蛋白偶联物(CIP-BSA),用紫外扫描、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法鉴定偶联物。利用制备的CIP-BSA偶联物作为免疫原,免疫Balb/c小鼠,用杂交瘤技术制备环丙沙星单克隆抗体,并对其效价、亲和力和特异性进行鉴定。结果表明:CIP-BSA的偶联比为30:1。经间接酶联免疫吸附(ELISA)法筛选,共筛出两株敏感、特异的杂交瘤细胞(JY-1、JY-2)。这两株细胞的腹水经纯化后的抗体效价分别为5.12×106、2.56×106,亲和力常数分别为2.88×109、2.46×109L/mol,两抗体均为IgG1a,取亲和力高的JY-1进行竞争抑制测定,IC50值可以达到9.83ng/mL。该单抗仅与恩诺沙星存在交叉反应,交叉反应率为95%。通过验证得到的JY-1细胞株分泌的单克隆抗体可用于水产品中环丙沙星残留的快速检测。
译  名：
Preparation and Identification of Monoclonal Antibody against Ciprofloxacin
作  者：
YANG Xian-le1,HUANG Xuan-yun1,HU Kun1,FANG Wei1,WANG Lian2,JIANG You-sheng1,*(1.National Pathogen Collection for Aquatic Animal,Shanghai Ocean University,Shanghai 201306,China ;2.Sichuan Provincial Fisheries School,Chengdu 401520,China)
关键词：
ciprofloxacin;hapten;antibody;immunity
摘  要：
The ciprofloxacin-bovine serum albumin conjugate(CIP-BSA) was prepared by a modified carbodiimide method,and identified by ultraviolet spectrometry and polyacrylamide gel electrophoresis(SDS-PAGE).Then CIP-BSA was used as immunogen to immunize BALB/c mice,inducing the secretion of antibodies against hapten CIP.The monoclonal antibodies(Mabs) against CIP were obtained by hybridoma technology and their titer,affinity and specificity were identified.The results showed that CIP-BSA coupling ratio was 30:1,and two Mabs secreted by cell lines JY-1 or JY-2 belonging to IgG1a were screened by the indirect ELISA method.The titers of purified JY-1 and JY-2 Mabs were 5.12 × 106 and 2.56 × 106,respectively;both the Mabs showed high affinity to CIP with affinity coefficients of 2.88 × 109 and 2.46 × 109 L/mol respectively.A competitive inhibition curve of the JY-1 Mab was developed,and the IC50 was 9.83 ng/mL.The JY-1 Mab exhibited high cross-reactivity(95%) with enrofloxacin.These results suggested that the JY-1 Mab was useful for determination of CIP residues in aqua-products.

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