Generation And Immunological Traits Of The Monoclonal Antibody Against Ciprofloxacin

The ciprofloxacin (CPFX) belongs to the fluoroquinolone (FQNs) group. As the second antibacterial agent of the FQNs group that was permeated to sale. It can efficiently damage the bacteria of Gram-positiv through inhibiting the DNA helicase. It has the advantages of broad-spectrum of antibacterial, low toxin, broadly spread in the body. So it was applied widely especially in the clinic of animals and production of farming, Use to treat digestive infections in farming industries as feed additives.The wide spread use of antibiotics in food-producing animals has caused public concern due to the transfer of antibiotic-resistant bacteria to man, It has the latent cause cancer and effect the health of human. In many countries, withdrawal period for CPFX and maximum residue limits (MRLs) in the food-producing animals have been set. So the CPFX detecting in tissues has become a necessary content. However, it demand we build a quickly detection for the ciprofloxacin residue.The objective of the paper is generate monoclonal antibodies (mAb) against CPFX and identify its immunological traits. In order to provide the technology support for detection blocking ELISA kit of CPFX residue in livestock products The CPFX was conjugated to carrier protein BSA and OVA by EDC to obtain Immunogen and coating antigen. The ultraviolet scanning and SDS-PAGE were used to identify CPFX artificial antigen. The results of scanning and SDS-PAGE showed that artificial antigen was synthesized successfully. Then immunized BALB/c mice with immunogen. Five mice was indirect ELISA titer against CPFX were all above 1:105 and the IC50 of No. 4 mice was the lowest (17.21ug/L). The hybridoma lines that secrete monoclonal antibody against CPFX (CPFX mAb) were established through cell fusion and indirect ELISA and blocking ELISA. One hybridoma cell line of 3D2 was screened for specificity to CPFX after cell fusing. CPFX mAb was generated by inducing ascites in mice. The immunological characteristics such as Karyotype, titer, sensitivity and specificity of the mAb were characterized. The indirect ELISA titer of the mAb was 1:1.28×103 in supernatant, 1:1.6×105 in ascites, The mAb of 3D2 showed high sensitivity with an IC50 of 9.95 ug/L to CPFX, The rate of cross reaction to Enrofloxacin could reach to 128.39%. There were low reactions to other fluoroquinolones antibiotics.The high-titer, sensitive and specific CPFX mAb has been generated and laid a solid foundation for the detection of ELISA kit of CPFX residue.