| Ceftiofur is cephalosporin used on veterinarian's clinic at first, because it is used extensively day by day in veterinarian, its residue in animal food has already aroused great concern from people gradually. To development of ELISA kit for rapid screening the residues of Ceftiofur,we prepared the monoclonal antibodies against Ceftiofur and developed the ELISA method.and 5 hybridomas secreting antibodies against ceftiofur were selected and cloned.A Ci-Elisa method for detecting ceftiofur was established. We also established detection method of ceftiofur by high-performance liquid chromatography at the same time.The immunogen and coating antigen were synthesized by carbodiimide method and isobutylchoroformate .The artificial antigens were identified by the Fecl3 colorimetry, UV absorbance and Infrared Spectrum method.The results demonstrated that the three kinds of artificial antigens were synthesized successfully.Four female BALB/c mice aged 4-6 weeks old were immunized intraperitoneally with the CFA adjuvant or IFA adjuvant in different dosages.All the antiserum titers were up to l:100000,and the free ceftiofur could compete with the coatings for the antibody.so there were specific anti-ceftiofur antibodies in the sera.Utilize cell's integration technology, merge the spleen cell of the little mouse and SP2/0 bone myeloma cell by 50% of the PEG, adopt indirect ELSIA law and compete for ELSIA law indirectly, utilize and dilute the clone technology of the law limitedly, clone and screening through 4 times , screen to 5 hybridization tumour cell one that can secrete the purpose antibody steadily s altogether. These hybridoma cell lines secreting anti-cef McAbs were obtained,these hybridomas were prepared ascites of the mice, and the titers of the McAbs in the ascetic fluid ranged from 10-4 to 10-6The subtype of 3B3 monoclonal antibody was IgM.A method of enzyme-linked) immunoassay was developed,and the 50% inhibition concentration of control... |
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