新霉素免疫亲和柱的制备及其性能鉴定 - 食品安全质量检测学报.pdf

7 3 Vol. 7 No. 3 2016 3 Journal of Food Safety and Quality Mar. , 2016 郑 琦, 曹立民, 郑洪伟, 林 洪, 隋建新* (, 266003) 摘 要: 目的 方法 A , sepharose 4B , ELISA ; , (HPLC) ; HPLC 结果 5.67 , 2.02×108 L/mol; 4 ℃6 h ; HPLC 0.2 mol/L (0.5 mol/L NaCl, pH 2.3), 104.57%±0.03%(n=6); 5 , 85%; 0.02%NaN3 PBS , 110 d 800 ng 75%; 50050007500 μg /kg , 63.29%±5.30% 80.15%±6.27%83.87%±0.88%; 2~4 h 结论 , 关键词: ; ; Preparation and characterization of an immunoaffinity column for the selective extraction of neomycin * ZHENG Qi, CAO Li-Min, ZHENG Hong-Wei, LIN Hong, SUI Jian-Xin (College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China) ABSTRACT: Objective To prepare and characterize an immune affinity column for the selective extraction of neomycin. Methods The immune affinity column of neomycin (NEO) was prepared by coupling CNBr-activated Sepharose-4B with the purified anti-NEO polyclonal antibody. An indirect competitive ELISA method was established to detect the specificity, affinity constant and titer of antibody. The changes of protein concentrations were determined by Coomassie brilliant blue method. HPLC method was used to explore the optimal elute conditio