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Development and validation of an immunochromatographic assay for rapid multi-residues detection of cephem antibiotics in animal-origined food author:xiehuiling Penems and cephems’ nucear structure Three antigenic determinants when the beta-lactam ring open ,these are the structural formula Beta-lactam antibiotics are able to generate several antigen determinants that are recognized by the immune system. That are common structure of nucleus and the acyl-side chain structure and a new antigenic determinant(NAD).in which the NAD formed by the conjugation of beta-lactam and carrier protein.the beta-lactam ring is easy to open when the hapten-protein conjugation is prepared by alkaline method. Let me introduce my work having finished and unfinished. Polyclonal Antibodies were produced in several ways. Group-specific antibody was raised by coupling 7-ACA/cephalexin to keyhole limpet hemocyanin (KLH) using glutaraldehyde as cross-linker. Specific antibody was raised by coupling cephadroxil to KLH using carbodiimide-mediated coupling method. Another specific antibody was raised by coupling ceftiofur to KLH using a method without any cross-linker or mediator molecule(physiological binding). Work have finished two rabbits were immunized with one kind immunogen. Now four polyclonal antibodies have been obtained,two of them were group-specific antibody,other two were specific antibody. Work I am doing Development of the antibody titer was monitored by an indirect enzyme-linked immunosorbent assay (ELISA),titer value is about 100000.the inhabition rate(50% binding inhabition concerntration)of cephalexin and cephadroxil were respectively 1-1.5 ng/ml and 1.5-2 ng/ml . Now The inhabition rate examination is being optimized Work I am doing A rapid colloidal gold assay was developing for the detection of cephems antibiotics in food of animal-origined. the polyclonal antibodies aginst cephalosporins was conjugated to colloidal gold
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