环丙沙星单克隆抗体的制备和性质鉴定  

Preparation and Characterization of Anti-ciprofloxacin Monoclonal Antibody

作　　者：顾黎[1,2] 孔蕴[1] 刘围[3] 郜海涛[1] 郗日沫[3] 王鹏[1,2] 

机构地区：[1]山东大学生命科学学院微生物技术国家重点实验室,济南250100 [2]国家糖工程技术研究中心,济南250100 [3]山东大学化学与化工学院,济南250100

出　　处：《科技通报》2010年第1期72-76,共5页Bulletin of Science and Technology

基　　金：山东省优秀中青年科学家科研奖励基金(2007BS02001);国家973项目(2007CB914800)资助

摘　　要：为了制备抗环丙沙星单克隆抗体,首先采用碳二亚胺法合成了环丙沙星与载体蛋白偶联的免疫抗原和包被抗原,免疫BALB/c小鼠,取效价最高的小鼠脾细胞与骨髓瘤细胞SP2/0融合,HAT培养基筛选,有限稀释法克隆化,获得1株稳定分泌抗环丙沙星单克隆抗体的杂交瘤细胞株。检测结果显示,杂交瘤细胞染色体数目平均102条,培养上清的效价为1∶1.28×103,单抗腹水效价达1∶1.024×106,IC50为68.31 ng/mL,属于IgM型,轻链kappa,CI-ELISA法显示单抗对环丙沙星有高度的选择性,提示获得的单抗可望用于环丙沙星残留物的免疫化学检测。For the aim of preparing anti-Ciprofloxacin monoclonal antibody, the immunogen CIP-cBSA and coating antigen CIP-cOVA were synthesized by EDC-method. Ultraviolate was used to identify the above artificial antigens. BALB/cmice were immunized by CIP-cBSA. Spleen cells of the mice with highest titer were fused with SP2/0 cells ,after selection with HAT medium and limiting colonization,one hybridoma cell line stable secreting McAb was established. Main characteristics of the acquired McAb were identified. The results showed that the hybridoma cells had around 102 chromosomes,the titer of the hybridoma cell supernatant and aseites were 1：1.28×10^3 and 1：1.024×10^6 respectively, determined by indirect ELISA. The IC50 was 68.31 ng/ml,and the isotype of McAb in aseites was IgM/κ. The specificity determined by CI-ELISA showed that the McAb had little cross reactivity to other fluoroquinolones antibiotics,except Enrofloxacin and Danofloxacin mesylate. The prepared CIP McAb was high-titer, sensitive and specific ,and then hopeful to be further developed to detect CIP redidues in animal foods.

关 键 词：环丙沙星 药物残留 单克隆抗体 

分 类 号：Q819[生物学—生物工程]