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Olaquindox (Olaquindox, OLA) is the chemical synthesis of quinoxaline broad-spectrum antimicrobial drugs, anabolic role to promote animal growth, improve feed conversion ratio and lean, can also be used for the prevention and treatment of bacterial infectious diseases. Production practice, olaquindox not regulate the use of not only directly harm the health of the animal's body, can also cause great harm to human health through the food chain. With the development of science and technology and national attention, the detection method has evolved. There electrochemical detection methods of analysis, spectrometry, chromatography, liquid chromatography - mass spectrometry, immunoassay technology. In order to establish a rapid, sensitive the olaquindox residual immunological analysis method, this study prepared olaquindox monoclonal antibody analysis olaquindox immunological characteristics based on the use of monoclonal antibody technology and assembly indirect competitive ELISA kit. The main findings are as follows: 1. Of olaquindox artificial antigen preparation and identification of succinic anhydride synthesis of derivatives the olaquindox - half succinate (OLA-HS); activation ester method synthetic full antigen OLA-BSA OLA-OVA ultraviolet spectrophotometry spectrophotometry (UV), gel electrophoresis (SDS-PAGE) identification of indirect ELISA; OLA-BSA immunized BALB / c mice, the antiserum titer, indirect competitive ELISA assay sensitivity and specificity. UV scan plots show the maximum absorption peak of the synthesis of artificial antigens offset occurred prove successfully coupled, and measuring the OLA-BSA, in OLA-OVA OLA-HS and BSA, OVA molecule binding ratios were 17.1:1 and 12.4:1; SDS-PAGE results showed that the the BSA swimming speed is greater than the OLA-BSA, the OLA-BSA molecular weight greater than BSA, further illustrate the success of OLA-HS and the BSA conjugate. # 3 mice antisera titer highest OLA the 50% inhibitory concentration (IC50) 58.923 ng / mL, was 1.841% cross-reactivity with carbadox, no cross-reaction with other drugs. High prices obtained through experiments, sensitive, and specific polyclonal antibody (pAb). Olaquindox of monoclonal antibodies and immunological characteristics identified indirect ELISA and indirect competitive ELISA select cell fusion spare mouse; hybridoma technology to prepare the olaquindox monoclonal antibody (OLAmAb), and its potency, affinity, sensitivity specific subtypes were identified; vivo inducible ascites method large scale preparation of monoclonal antibodies. Screened 2B1, 4F5, 4E12, 5B5, 5E5 the five sensitivity and specificity of hybridoma; identification of monoclonal antibody subtypes are IgG1; indirect ELISA titer in cell culture supernatant in 1:3.0 × 102 1 5 between sup>: 1.28 × 10 3 , OLAmAb affinity constant Ka of 3.75 x 10 < sup> 10 L / moL, OLA IC50 of 1.66ng/mL; cross-reactivity with carbadox rate of 5.19%, and no cross-reaction with other drugs. High titer obtained by experiment, sensitive, specific OLAmAb, immunologic tests that can be used for the detection of residues of animal food in OLA. 3 Olaquindox residual immunological rapid method for detection of the enzyme-linked immunosorbent assay principle, the application of OLAmAb developed OLA residual indirect competitive ELISA for rapid detection reagent box (OLA-Kit), the OLA-Kit standard curve is S-shaped, in line with 4 parameter logit curve fitting, the minimum detection limit 0.126μg / L, the IC50 1.66ng/mL, average intra-and inter-assay coefficient of variation of less than lt; 15%. Pork liver, pork, add 2,10,50,100 ng / mL the OLA standards, the average recoveries were 77.6% and 79.68% respectively. OLA-Kit performance is stable and can be stored for 6 months at 4 ℃.
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