Xu Bojie, Zhang Xiangyu, Wang Kai, Xue Xianle, Hongsibsong Surat, Morisseau Christophe, Li Qing X, Hammock Bruce D, Xu Ting
Beijing Key Laboratory of Biodiversity and Organic Farming, College of Resources and Environmental Sciences, China Agricultural University, Beijing, 100193, China.
Research Institute for Health Science, Chiang Mai University, Chiang Mai, 50200, Thailand.
Talanta. 2025 Sep 1;292:127980. doi: 10.1016/j.talanta.2025.127980. Epub 2025 Mar 18.
Triclosan (TCS), a broad-spectrum antimicrobial agent in pharmaceuticals and personal care products, necessitates environmental monitoring due to its antimicrobial properties and widespread in the environment. In this study, two anti-TCS single-domain antibodies (i.e., nanobodies, Nbs), T1 and T2, were isolated from a phage-displayed Nb library derived from a camel immunized with a mixture of TCS immunogens. The T1-based enzyme-linked immunosorbent assay (ELISA) exhibited a better sensitivity to TCS than the T2-base ELISA. Motivation at enhancing specificity, sensitivity, and stability of Nb-based immunoassays promoted exploring use of a bivalent strategy to enhance performance. The bivalent Nb T1-T1 was tandemized via a linker-(GGGGS)3- between. The thermal stability of the bivalent Nb was improved in comparison with that of a monovalent Nb. The sensitivity of T1-T1-based ELISA, with an IC value of 4.3 ng mL of TCS, was improved approximately 2-3 fold in comparison to those of T1-or T2-based ELISAs (8.5 and 14.6 ng mL, respectively). The average recovery of TCS from water samples measured with T1-T1-based ELISA was in the range of 99-125 %, which correlated well with that measured by a high-performance liquid chromatography (HPLC) method (R > 0.99). TCS in river water samples was detected by the resultant ELISA and an HPLC method, showing a satisfactory correlation.
三氯生（TCS）是药品和个人护理产品中的一种广谱抗菌剂，因其抗菌特性且在环境中广泛存在，所以需要对其进行环境监测。在本研究中，从用TCS免疫原混合物免疫的骆驼所构建的噬菌体展示纳米抗体（Nb）文库中分离出两种抗TCS单域抗体（即纳米抗体，Nb），T1和T2。基于T1的酶联免疫吸附测定（ELISA）对TCS的敏感性优于基于T2的ELISA。为提高基于Nb的免疫测定的特异性、敏感性和稳定性，促使人们探索使用二价策略来提升性能。二价Nb T1-T1通过连接子-(GGGGS)3-进行串联。与单价Nb相比，二价Nb的热稳定性得到了提高。基于T1-T1的ELISA对TCS的敏感性提高，其IC值为4.3 ng/mL TCS，与基于T1或T2的ELISA（分别为8.5和14.6 ng/mL）相比提高了约2-3倍。用基于T1-T1的ELISA测定水样中TCS的平均回收率在99%-125%范围内，与通过高效液相色谱（HPLC）法测得的回收率相关性良好（R>0.99）。用所得ELISA和HPLC法检测河水样中的TCS，显示出令人满意的相关性。
