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

发布时间：2018-01-30 19:16

本文关键词： 拉沙里菌素半抗原单克隆抗体间接竞争ELISA　出处：《中国畜牧兽医》2017年10期 　论文类型：期刊论文

【摘要】：为制备拉沙里菌素(LAS)单克隆抗体,建立针对LAS的间接竞争ELISA(Ci-ELISA)方法,试验采用了活性酯法将LAS分别与BSA和OVA偶联成LAS-BSA和LAS-OVA作为免疫原和检测原,6次免疫后,取小鼠脾细胞与骨髓瘤细胞融合,最终筛选出一株能稳定分泌抗LAS单克隆抗体的杂交瘤细胞株C11。经鉴定,C11的抗体类型为Ig G1,轻链为κ链;所制腹水效价为1∶16 000,与莫能菌素钠、盐霉素钠、马杜霉素胺、头孢噻吩钠和硫酸链霉素均无交叉反应。用此单克隆抗体建立LAS Ci-ELISA检测方法显示,Ci-ELISA标准工作曲线为y=0.376x-0.2374(R2=0.9914),LAS在5~1 000 ng/m L时,线性关系良好,IC50为90.22 ng/m L,加标回收率为81.76%~102.41%,表明方法精确度好、灵敏度高。LAS单克隆抗体的制备和Ci-ELISA方法的建立为下一步试剂盒的研发奠定了基础。
[Abstract]:In order to prepare LAS-LAS monoclonal antibody, an indirect competitive ELISA-Ci-ELISA-ELISA-specific method for LAS was established. In this experiment, LAS was coupled with BSA and OVA to form LAS-BSA and LAS-OVA as immunogen respectively by using active ester method. Mouse spleen cells were fused with myeloma cells, and a hybridoma cell line C11, which could stably secrete monoclonal antibody to LAS, was selected. The antibody type of C11 was identified as Ig G1. The light chain is 魏 chain; The titer of ascitic fluid was 1:16, with monensin sodium, salinomycin sodium and maduromycin amine. There was no cross reaction between cefthiophene sodium and streptomycin sulfate. LAS Ci-ELISA assay was established with this monoclonal antibody. The standard working curve of Ci-ELISA was 0.376x-0.2374 R29914L, and the linear relationship was good when the value of Las was 50.376x-0.2374 ng/m / L. The IC50 was 90.22 ng/m / L and the recovery rate was 81.76 / 102.41, which indicated that the method was accurate. The preparation of highly sensitive. Las monoclonal antibody and the establishment of Ci-ELISA method laid a foundation for the further development of the kit.
【作者单位】：扬州大学兽医学院;江苏省动物重要疫病与人兽共患病防控协同创新中心;
【基金】：牛羊碳水化合物、蛋白质与能量代谢失衡防控技术研究(2016YFD0501206)
【分类号】：S859.84
【正文快照】： 拉沙里菌素(lasalocid,LAS),又名那塞罗,是由拉沙里链霉菌发酵产生一种的抗生素[1-2],与莫能菌素、盐霉素同属聚醚类离子载体抗生素。LAS主要用于提高动物生长性能,改善反刍动物的饲料利用率[1-3],预防畜禽球虫感染[4]。LAS摄入后会分布在肝脏、肌肉、脂肪、心脏、皮肤、胸腺

【相似文献】