Broad-spectrum detection of benzimidazoles with lateral flow immunoassay: A computational chemistry-assisted hapten design strategy and explore of molecular recognition mechanism

• Published in: Journal of hazardous materials 2024-12, Vol.480, p.135986, Article 135986
• Main Authors: Yin, Jiaqi, Yu, Sha, Mei, Xi, Chen, Zongyou, Peng, Juan, Lai, Weihua
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; Benzimidazoles; Benzimidazoles - chemistry; Broad-Spectrum; Computational Chemistry; Food Contamination - analysis; Hapten design; Haptens - chemistry; Haptens - immunology; Immunoassay - methods; Limit of Detection; Mice, Inbred BALB C; Monoclonal antibody; Recognition mechanism
• ISSN: 0304-3894; 1873-3336; 1873-3336
• DOI: 10.1016/j.jhazmat.2024.135986

Benzimidazoles (BMZs) are a class of veterinary drugs with a benzimidazole ring, the abuse of which poses a serious threat to ecological balance and human health. Consequently, the development of broad-spectrum antibodies and rapid assays are crucial for detecting BMZs in food samples. Herein, we scientifically designed three hapten structures, predicted the availability of the hapten with computational chemistry, and subsequently verified the broad-spectrum with immunological experiments. A broad-spectrum monoclonal antibody (6F10) was prepared based on the predicted hapten-2. Molecular recognition studies illustrated intricate interactions between mAb 6F10 binding to BMZs attributed to halogen bonds and π-π/π-alkyl interactions, revealing key amino acid sites and demonstrating the reliability of the hapten prediction strategies. Finally, a broad-spectrum, rapid, and sensitive lateral flow immunoassay based on aggregation-induced emission microspheres with high fluorescence intensity was established. The LOD values of the proposed method for eight kinds of BMZs were 0.027, 0.032, 0.058, 0.091, 0.087, 0.246, 0.369, and 0.311 ng mL−1, respectively. In this work, a hapten prediction strategy based on a computational chemistry method effectively guided the preparation of antibodies for broad-spectrum recognition of BMZs, and the molecular recognition studies verified the interaction of mAb 6F10 with BMZs, enabling broad-spectrum and sensitive detection of BMZs in milk. [Display omitted] ●Prediction the availability of haptens structures with computational chemistry.●A broad-spectrum mAb 6F10 to benzimidazoles was prepared.●The recognition mechanism of mAb 6F10 was investigated by molecular simulation.●A broad-spectrum and sensitive LFIA for detection of BMZs in milk samples.●The proposed LFIA is a promising candidate for environmental monitoring.