Development of a single-chain variable fragment-alkaline phosphatase fusion protein-based direct competitive enzyme-linked immunosorbent assay for furaltadone metabolite in ctenopharyngodon idellus

• DOI: 10.1080/09540105.2021.1957781
• Publication Date: 2021-08-01T20:34:26Z
• Authors: Xinzhu Chen; Huaying Du; Yanping Hong; Yubo Wang; Mei Shu; Dan Wang; Peng Dai; Weijie Deng; Jianhua Xiong; Wuying Yang
• Keywords: Agriculture (General); furaltadone metabolite amoz; single-chain variable fragment antibody; direct competitive enzyme-linked immunosorbent assay; 04 agricultural and veterinary sciences; Immunologic diseases. Allergy; RC581-607; 0405 other agricultural sciences; alkaline phosphatase; scfv-ap fusion protein; S1-972

To determine furaltadone metabolites 5-methylmorpholino-3-amino-2-oxazolidinone (AMOZ) residual levels in ctenopharyngodon idellus, direct competitive enzyme-linked immunosorbent assay (dcELISA) was developed based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein. Firstly, VH and VL gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against AMOZ derivative, then the productive genes assembled to scFv engineered produce scFv-AP The protein further identified as bifunctional reagent for immunoassay, sensitive one-step dcELISA derivative with half-maximal inhibitory concentration (IC50) limit of detection (LOD) 10.62 4.83 ng/mL, respectively. recovery values spiked fish samples determined by 71.31%∼82.88%. results suggested that prepared this work can be used rapid convenient immunoassays detect residues samples.