Abstract:

An enzyme-linked immunoabsorbence (ELISA) test kit was developed for enrofloxacin (ENR) detection. ENR-MBSA and ENR-OVA antigens were prepared by EDC-NHS method. BALB/C mice were immunized with ENR-M-BSA for monoclonal antibody generation. A specific monoclonal antibody was screened using methylcellulose-based semi-solid medium and analyzed using indirect competition ELISA. The molar coupling ratios of ENR-conjugated BSA and ENR-conjugated OVA were calculated to be 15:1 and 7.2:1, respectively. The specific monoclonal antibody 3E9 was obtained and the titer of purified 3E9 antibody reached 107 according to ELISA detection, and belonged to the family of IgG1 according to the detection using an antibody typing kit. The ELISA detection of ENR using the antibody exhibited a sensitivity of 1 ng/mL, a linear range of 1 to 100 ng/mL and a half inhibitory concentration (IC50) of 10 ng/mL. The cross-reactivity between the ELISA detection kit developed and four analogues was less than 1%. These results indicate that the monoclonal antibody has a promising potential in the development of ELISA test kits for ENR detection.

Key words: enrofloxacin (ENR), ELISA, monoclonal antibody