5-甲基水杨酸的免疫原和抗体的制备与应用
Preparation and Application of Immunogen and Antibody of 5-Methylsalicylic Acid
伍华琛 1庄惠生1
作者信息
- 1. 上海交通大学环境科学与工程学院,上海200240;上海交通大学环境监测评价研究所,上海200240
- 折叠
摘要
通过活性酯与碳二亚胺联合法或混合酸酐法将5-甲基水杨酸与牛血清白蛋白偶联制得偶联比分别为14.6和3.4的免疫原(简称1和2).用1和2分别免疫雄性新西兰大白兔得到两种多克隆抗体(简称3和4).用间接竞争ELISA测得3和4的抗体效价分别为1:256000与1∶128 000.用3建立了测定5-甲基水杨酸的间接竞争ELISA法.该方法的IC50为149.04 ng·mL-1,最低检测限为4.19 ng·mL-1;3与苯酚、对甲酚和邻氨基四甲基苯酚的交叉反应率均＜0.1％.
Abstract
Two immunogens(1 and 2) were prepared by coupling of 5-methylsalicylic acid with BSA through a combination method of active ester and EDC or a mixed-anhydride method.The coupling ratios of 1 and 2 are 14.6 and 3.4,respectively.Two antibodies (3 and 4) were obtained by the stimulating male New Zealand rabbit with 1 or 2.3 and 4 were characterized by i-Elisa methods.The results showed that the titer of 3 and 4 were 1:256 000 and 1:128 000.An i-Elisa method for 5-methylsalicylic was established with 3,and the results indicated that IC50 was 149.04 ng · mL-1 and LOD was 4.19 ng· mL-1.The cross-reactivity rates of 3 with 2-amino-p-cresol,phenol and cresol were all less than 0.1％.
关键词
5-甲基水杨酸/偶联比/间接竞争ELISA/免疫原/抗体/制备Key words
5-methylsalicylie acid/coupling ratio/indirect competitive ELISA/immunogen/antibody/preparation引用本文复制引用
基金项目
出版年
2013
NETL
NSTL
维普
万方数据