Nanobody-Based Enzyme Immunoassay for Aflatoxin in Agro-Products with High Tolerance to Cosolvent Methanol

文献类型： 外文期刊

第一作者： He, Ting

作者： He, Ting;Wang, Yanru;Li, Peiwu;Zhang, Qi;Lei, Jiawen;Zhang, Zhaowei;Ding, Xiaoxia;Zhou, Haiyan;Zhang, Wen;He, Ting;Wang, Yanru;Li, Peiwu;Lei, Jiawen;Zhang, Zhaowei;Ding, Xiaoxia;Zhang, Wen;Li, Peiwu;Zhang, Qi;Zhang, Zhaowei;He, Ting;Wang, Yanru;Li, Peiwu;Zhang, Qi;Lei, Jiawen;Zhang, Zhaowei;Ding, Xiaoxia;Zhou, Haiyan;Li, Peiwu;Ding, Xiaoxia;Zhou, Haiyan;Zhang, Wen

作者机构：

期刊名称：ANALYTICAL CHEMISTRY （ 影响因子：6.986； 五年影响因子：6.755 ）

ISSN： 0003-2700

年卷期： 2014 年 86 卷 17 期

页码：

收录情况： SCI

摘要： A phage-displayed library of variable domain of heavy chain of the heavy chain antibody (VHH) or nanobody (Nb) was constructed after immunizing an alpaca with aflatoxin B1 (AFB(1)) conjugated with bovine serum albumin (AFB(1)-BSA). Two AFB(1)-specific nanobodies were selected. The obtained nanobodies were compared to an aflatoxin-specific monoclonal antibody B5 with respect to stability under organic solvents and high temperature. The two nanobodies could bind antigen specifically after exposure to temperatures as high as 95 degrees C. Besides, the nanobodies showed better or similar tolerance to organic solvents. A competitive ELISA with nanobody Nb26 was developed for the analysis of AFB(1), exhibiting an IC50 value of 0.754 ng/mL (2.4 mu M), linear range from 0.117 to 5.676 ng/mL. Due to the high tolerance to methanol, sample extracts were analyzed by nanobody-based ELISA without dilution. The recovery from spiked peanut, rice, corn and feedstuff ranged from 80 to 115%. In conclusion, the isolated nanobodies are excellent candidates for immunoassay application in aflatoxin determination.

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