Hapten Synthesis and Monoclonal Antibody Preparation for Simultaneous Detection of Albendazole and Its Metabolites in Animal-Origin Food

• Published in: Foods Vol. 10; no. 12; p. 3106
• Main Authors: Shao, Shibei, Zhou, Xuping, Dou, Leina, Bai, Yuchen, Mi, Jiafei, Yu, Wenbo, Zhang, Suxia, Wang, Zhanhui, Wen, Kai
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 14.12.2021; MDPI
• Subjects: Albendazole; Anthelmintic agents; antibody; Antigens; Antiparasitic agents; Benzimidazoles; Cell culture; Chromatography; Coefficient of variation; Computational chemistry; Computer applications; Conformation; Drug resistance; Electric fields; Electrostatic properties; Enzyme-linked immunosorbent assay; Food safety; Food science; hapten design; Hydrophobicity; Immune system; Immunoassay; Laboratory animals; Mass spectrometry; Metabolites; Monoclonal antibodies; Recognition; Scientific imaging; Beijing China; United States > US; China; hapten design; antibody; computational chemistry; albendazole; metabolites; immunoassay
• ISSN: 2304-8158; 2304-8158
• DOI: 10.3390/foods10123106

Albendazole (ABZ) is one of the benzimidazole anthelmintics, and the overuse of ABZ in breeding industry can lead to drug resistance and a variety of toxic effects in humans. Since the residue markers of ABZ are the sum of ABZ and three metabolites (collectively referred to as ABZs), albendazole-sulfone (ABZSO ), albendazole-sulfoxide (ABZSO), and albendazole-2-amino-sulfone (ABZNH SO ), an antibody able to simultaneously recognize ABZs with high affinity is in urgent need to develop immunoassay for screening purpose. In this work, an unreported hapten, 5-(propylthio)-1H-benzo[d]imidazol-2-amine, was designed and synthesized, which maximally exposed the characteristic sulfanyl group of ABZ to the animal immune system to induce expected antibody. One monoclonal antibody (Mab) that can simultaneously detect ABZs was obtained with IC values of 0.20, 0.26, 0.77, and 10.5 μg/L for ABZ, ABZSO , ABZSO, and ABZNH SO in ic-ELISA under optimized conditions respectively, which has been never achieved in previous reports. For insight into the recognition profiles of the Mab, we used computational chemistry method to parameterize cross-reactive molecules in aspects of conformation, electrostatic fields, and hydrophobicity, revealing that the hydrophobicity and conformation of characteristic group of molecules might be the key factors that together influence antibody recognition with analytes. Furthermore, the practicability of the developed ic-ELISA was verified by detecting ABZs in spiked milk, beef, and liver samples with recoveries of 60% to 108.8% and coefficient of variation (CV) of 1.0% to 15.9%.