微囊藻毒素-LR免疫亲和层析柱的研制和应用  

Preparation and Application of Affinity Chromatography Column for Microcystin-LR

作　　者：肖付刚[1] 赵晓联[2] 汤坚[1] 顾小红[1] 张敬平[3] 钮伟民[3] 

机构地区：[1]江南大学食品科学与技术国家重点实验室,江南大学分析测试中心,无锡214122 [2]江苏省微生物研究所有限公司,无锡214063 [3]无锡市疾病预防控制中心,无锡214023

出　　处：《分析化学》2008年第1期99-102,共4页Chinese Journal of Analytical Chemistry

基　　金：国家卫生部资助课题(No.WKJ2006-2-10)

摘　　要：用CNBr-activated Sepharose4B和微囊藻毒素-LR的单克隆抗体制备了免疫亲和层析柱,测得抗体偶联率在75.7%～94.1%之间。制得的免疫亲和层析柱最大柱容量在76～95ng之间,柱空白为0,回收率在90.8%～105.1%之间。柱子再生重复使用6次后,回收率不低于75%。建立了免疫亲和层析柱-高效液相色谱测定水样中的微囊藻毒素-LR的方法。该法检出限为5ng/L;线性定量范围为10～500ng/L。实验结果显示,免疫亲和层析柱特异性好,一次净化能除去绝大部分干扰物,净化效果明显优于现有的固相萃取柱。Immunoaffinity chromatography column was prepared with CNBr-activated Sepharose 4B and monoclonal antibody of microcystin-LR and its coupling rate is 75.7% -94. 1%. The column load for microcystin- LR is 76-95 ng and blank is 0, the recovery is 90. 8% - 105.1%. This column can be regenerated 6 times and the recovering rate is no less than 75%. The method for trace analysis of microcystin-LR in water using immunoaffinity chromatography column and high performance liquid chromatograph was established, its limit of detection is 5 ng/L and linear range is 10 -500 ng/L. The result indicates that microcystin-LR can be recognized especially by immunoaffinity chromatography column. Most interference can be removed through once purification, the effect is better than that with solid phase extraction column.

关 键 词：微囊藻毒素-LR 免疫亲和层析 单克隆抗体 高效液相色谱 

分 类 号：R392[医药卫生—免疫学]