氯霉素多克隆抗体的制备及免疫活性研究

被引:10
作者
黄雅丽
程敬丽
桂文君
朱国念
机构
[1] 浙江大学农药与环境毒理研究所
关键词
氯霉素; 半抗原合成; 人工抗原; 抗体; ELISA;
D O I
暂无
中图分类号
S852.4 [家畜免疫学];
学科分类号
090602 ;
摘要
为建立氯霉素的免疫分析方法,由氯霉素原药与丁二酸酐反应合成了氯霉素两种半抗原(CAP-HS1、CAP-HS2),分别纯化后经质谱法(MS)分析鉴定结构.纯化的半抗原与牛血清蛋白(BSA)以碳二亚胺法偶联得到人工抗原.将免疫抗原分别免疫兔子获得了相应抗体.当包被抗原的浓度为10μg?mL-1时,抗CAP-HS1和CAP-HS2的冻干粉效价分别为8.39×106和6.55×104(OD490 nm=1.0).研究了抗原抗体反应的亲和力和竞争抑制能力,其中抗CAP-HS1抗体适合免疫分析,检测线性范围为0.2~100 ng?mL-1,最低检测限IC20为0.239 ng?mL-1,抗CAP-HS1抗体对各类似物如甲砜霉素、氟甲砜霉素的交叉反应率分别为0.025%和0.166%,对其他类抗生素的交叉反应率<2×10-5.
引用
收藏
页码:180 / 185
页数:6
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