庆大霉素免疫亲和柱的制备及测定

Preparation of Immunoaffinity Column for Separation and Indrect ELISA Determination of Gentamycin

建立庆大霉素残留的间接竞争ELISA检测方法及其用于样品处理的免疫亲和柱的制备。结果表明:ELISA方法的IC50为3.8ng/ml,线性范围为0.1～25ng/ml(R2=0.99),检测限为0.1ng/ml;用该抗体做的免疫亲和柱使用条件进行了探索,表明当洗脱液为70%甲醇-磷酸盐缓冲液(7:3,V/V),体积为5ml时,可以达到最佳的洗脱;利用庆大霉素标准品建立了HPLC测定的标准曲线,测得吸附率均在90%以上,回收率均在60%以上,可以重复使用大约5次左右,说明该免疫亲和柱的灵敏性和可靠性。

Gentamycin residues were analyzed through indrect ELISA method,and immunoaffinity columns specific to gentamycin was prepared for its separation.The optimal ELISA for detecting gentamycin in swine tissues was studied.The IC50 for gentamycin was 3.8 ng/ml.The linear range was 0.1～25 ng/ml（R2=0.99） with a detection limit of 0.1 ng/ml.The immunoaffinity column was prepared using such antibody,and could be eluted with 5ml methanol/PBS（7：3,V/V） solution.The rate of adsroption to gentamycin was more than 90% and recovery more than 60%,could be repeatedly used for about 5 times.