Preparation of Monoclonal Antibody and Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay and Fluorescence-Linked Immunosorbent Assay for Detecting 3-Amino-5-methylmorpholino-2-oxazolidinone (AMOZ) in Edible Animal Tissue

Abstract

To monitor the illegal used of furaltadone, a highly sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and fluorescence-linked immunosorbent assay (FLISA) based on a monoclonal antibody (mAb) were developed for the detection of 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), the major metabolite of furaltadone in animal tissues. The highly specific mAb, which was very sensitive to a nitrophenyl derivative of AMOZ (2-NP-AMOZ) with IC50 values of 0.11 and 0.09 ng/mL for ic-ELISA and FLISA, respectively, was selected for the development of immunoassays. For both the ic-ELISA and FLISA for AMOZ-spiked experiments, acceptable recovery rates of 81.1–105.3% and coefficients of variation of 4.7–9.8% were obtained. In addition, results from both ic-ELISA and FLISA methods for spiked samples’ data showed excellent correlation coefficients ranging from 0.9652 to 0.9927. Meanwhile, the proposed ic-ELISA and FLISA for thirty spiked samples were confirmed by standard LC-MS/MS with high correlation coefficients of 0.9911 and 0.9921, respectively. These results suggest that the developed ic-ELISA and FLISA are valid and cost-effective tools for high-throughput monitoring methods for AMOZ residues in animal tissues.

Immunosensor of Nitrofuran Antibiotics and Their Metabolites in Animal-Derived Foods: A Review

“…At the same time, there are discrete capping agents and different functional groups on its surface, thus increasing the sensitivity of the method ( Biranje et al, 2021 ). Xie et al ( Xie et al, 2021 ) established both ic-ELISA and Fluorescence-Linked Immunosorbent Assay (FLISA) based on QDs to detect AMOZ. The IC 50 values of ic-ELISA and FLISA were 0.11 and 0.09 μg/L, respectively, the recovery rate is 81.1–105.3%, and the coefficient of variation was 4.7–9.8%.…”

Hierarchical 3D Snowflake-like Iron Diselenide: A Robust Electrocatalyst for Furaltadone Detection

“…19−21 Previous literature reported that FLD is highly unstable in vivo and can be rapidly metabolized in the forms of 3-amino-5morpholinomethyl-2-oxazolidinone, 1-aminohydantoin, 3amino-2-oxazolidinone, and semicarbazide. 22,23 It has the potential to bind to tissue proteins with high affinity and generate long-lasting protein conjugates. 24 FLD and its metabolites can cause potential carcinogenic, mutagenic, and teratogenic effects on all living organisms.…”

“…Previous literature reported that FLD is highly unstable in vivo and can be rapidly metabolized in the forms of 3-amino-5-morpholinomethyl-2-oxazolidinone, 1-aminohydantoin, 3-amino-2-oxazolidinone, and semicarbazide. , It has the potential to bind to tissue proteins with high affinity and generate long-lasting protein conjugates . FLD and its metabolites can cause potential carcinogenic, mutagenic, and teratogenic effects on all living organisms. , Due to the negative impact on humans and the environment, FLD has been strictly prohibited in the United States (Register, 2002), China (Regulation of Department of Agriculture of China, 2002), European Union (EU Council Regulation No.…”