Production and characterization of monoclonal antibody against the metabolite of furaltadone, 5-morpholino-3-amino-2-oxazolidone (AMOZ) , and establishment of colloidal gold immunochromatography for detecting

Object: The monoclonal antibody against 5-morpholino-3-amino-2-oxazolidone ( AMOZ) was prepared, and the corresponding colloidal gold immunochromatography method was established. Method: The hapten CP-AMOZ was synthesized by the reaction between AMOZ and 4-CBA and conjugated with BSA and OVA using activated ester method.Ascitic type antibody was prepared by hybridom cell lines which secret monoclonal antibody against CP-AMOZ by mean of immunizing BALB/c mice, cell fusion, ELISA screening and subcloning. Data including the titer of antibody, its IC 50 against CP-AMOZ and LOD were detected by ELISA analysis and determine its specificity. Competitive colloidal gold immunochromatographic dipstick was produced on this base, gold nanoparticles were coupled with monoclonal antibodies, CP-AMOZ-BSA and rabbit-anti-mouse polyclonal antibody were separately coated on nitrocellulose membrane as test line and control line to detect the limit of detection and specificity.Result: The titer of the mAbs was 1 ∶1.6 × 105 , the IC 50 value and LOD to CP-AMOZ were 0.86μg /L and 0.07μg/L respectively. The mAbs had little cross reactivity with other nitrofurans metabolites and derivates, showing high specificity to CP-AMOZ, and LOD of colloidal gold immunochromatographic dipstick was 5μg /L.Conclusion: The antigen of CP-AMOZ was successfully synthetized and the monoclonal antibody specific to CP-AMOZ was prepared, which help prepare the colloidal gold rapid detection dipstick of high sensitivity and specificity.