中国畜牧兽医2017，Vol.45Issue(10)：3049-3056,8.DOI:10.16431/j.cnki.1671-7236.2017.10.031
拉沙里菌素单克隆抗体的研制及间接竞争ELISA检测方法的建立
Preparation of Monoclonal Antibody Against Lasalocid and Deveopment of Indirect Competitive ELISA Detection Method
摘要
Abstract
To prepare monoclonal antibodies (MAb) against lasalocid (LAS) and establish an indirect competitive ELISA (Ci-ELISA) detection method,conjuaction of LAS-BSA and LAS-OVA were synthetized as the immunogen and coating antigen by using active ester method in this experiment.After 6 times of immunization,the spleen cells of mice and myeloma cells were fused.Finally,a hybridoma cell line that could stably secrete specific MAb against LAS was screened.The immunological subtype of the MAb was identified as IgG1,and its light chain was κ type.The titer of the ascites was 1 ∶ 16 000,which showed no cross activity with monensin sodium,salinomycin sodium,maduramycin,cefalotin sodium and streptomycin sulfate.Ci-ELISA method was established based on the MAb against LAS with the linear equation was y=0.376x-0.2374(R2 =0.9914),and the linear range was 5 to 1 000 ng/mL and the IC50 was 90.22 ng/mL.The method was used to detect LAS in spiked samples and the recovery rate was 81.76％ to 102.41％within the detection range,indicating that the method was successfully established with good accuracy and high sensitivity.The preparation of MAb against LAS and the establishment of Ci-ELISA method laid the foundation for the development of LAS detection kit.关键词
拉沙里菌素/半抗原/单克隆抗体/间接竞争ELISAKey words
lasalocid/hapten/monoclonal antibody/indirect competitive ELISA分类
农业科技引用本文复制引用
杨小康,张绘艳,顾建红,袁燕,卞建春,刘宗平,刘学忠..拉沙里菌素单克隆抗体的研制及间接竞争ELISA检测方法的建立[J].中国畜牧兽医,2017,45(10):3049-3056,8.基金项目
牛羊碳水化合物、蛋白质与能量代谢失衡防控技术研究(2016YFD0501206) （2016YFD0501206）
