Preparation of monoclonal antibody against chloramphenicol and establishment of direct competitive ELISA for determination of chloramphenicol
抗氯霉素单克隆抗体的制备及直接竞争ELISA方法的建立

Through immunization of BALB/c mice with synthesized CAP-ovalbumin (CAP-OVA), a hybridoma cell line, the 4C9 which could produce monoclonal antibody against chloramphenicol (CAP) and propagate stably, was established. With the myeloma cells, the ascites containing monclonal antibody (McAb) were prepared. The indirect competitive ELISA (ciELISA) test revealed the heavy chain and light chain of McAb were IgG1 and κ, respectively. The ELISA titer of ascites was 1:1×10~6. The percentages of cross-activity to thiamphenicol, florfenicol and other antibiotics were all less than 0.01%. A horseradish peroxidase (HRP)-labeled antibody was synthesized by the sodium periodate reaction, then a direct competitive enzyme-linked immunosorbent assay (cdELISA) was developed. Quantization of the CAP was linear from 0.1 to 100 ng·mL~(-1), and the hemi-inhibitory concentration (IC_(50)) was 5.81 ng·L~(-1). The recovery test showed that the detection limit for CAP was 0.1 ng·L~(-1) in cdELISA. Comparative test showed that detected sensibility of the monoclonal antibody was nearly equivalent to the commercial CAP ELISA kit.