Production Of Antibodies Against Sulfadiazine And Establishment Of Enzyme-Linked Immunosorbent Assay

SD-BSA and SD-Tg conjugants were prepared by N-(3-Dimethy-laminoproyl)-N'ethylcarbodiimide hydrochloride (EDC), glutaraldehyde and diazotation reaction method respectively. Polyclonal antibodies against SD were obtained by immunizing rabbits with SD-BSA. The affinity constant of the anti-SD polyclonal antibodies were 2.9×10~9L/mol.The cross reactivities with SMT and SQ were less than 0.2% and 0.01% respectively. Four hybridoms cell lines secreting monoclonal antibody against SD were obtained. The affinity constant of the anti-SD monoclonal antibody SD2D5 and SD4D11 were 4.0×10~9L/moland 1.4×10~9L/mol respectively. and the cross reactivities with SMT and SQ were less than 0. 01% respectively. Different modes of ELISA for the measurement of SD were established. A two step competitive enzyme-linked immunoassay (ELISA) was developed by using SD2D5 to measure the residues of (Sulfadiazine) SD in food. The sensitivity of the assay was 0.023ng/mL. The intra-assay and inter-assay CVs were 9.9%～14.8%, and 7. 8%～17.6% respectively. The correlation coefficient between measured and expected values was 0. 995 after serial dilutions of the samples with high concentration of SD.