微囊藻毒素-LR免疫亲和层析柱的研制和应用

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。

Preparation and Application of Affinity Chromatography Column for Microcystin-LR

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.