Preparation And Identification Of Monoclonal Antibody Against Enrofloxacin And Establishment Of Immunoassay Method For Enrofloxacin

Enrofloxacin(Enro) was a kind of Fluoroquinolone antibiotics which were usually used in clinical therapy of bacterial infection of livestock, but the residues of it in meat of domestic animals severely threatened the health of public. It is very important to establish a kind of credible, sensitive and quick assay. Enro was coupled with carrier protein bovine serum album(BSA) and ovalbumin (OVA) to prepare the immune antigen Enro-BSA and the detection antigen Enro-OVA by N-hydroxysuccinimide ester (NHS) method respectively. Conjugated antigens were analysized by the UV scanning and animal experiment. The results showed that the conjugated antigens were successfully obtained and could be used to prepare the monoclonal antibody against Enro with immune antigen Enro-BSA and detection antigen Enro-OVA . Mice were immunized with Enro-BSA for 4 times. Anti-Enro monoclonal antibodies were produced by conventional cell fusion technique. The spleen cells of immunized Blab/c mice were fused with SP2/0 myeloma cells using PEG, and selected with HAT and HT medium. Tow hybridoma cell lines of 3A8 and 5D7 were obtained using Enro-OVA by indirect ELISA, and cloned by limited dilution method for three times, whose titers of cultivate medium and ascites were 1:1600, 1:1000 and 1:8.0×10~4, 1 : 3.2×10~3 respectively. The subtype of them were both...