| This paper reported the preparation of CL-McAb and the development of the competitive enzyme-linked immunosorbent assay (ELISA) for the detection of clenbuterol residues.β2- agonist clenbuterol has been used as bronchiodilators in human medicine for over 30 years. Unfortunately, this agent has also been used illegally now as growth-promoter in livestock production. Its illegal use will lead to toxic effects after human consumption of meat products. The problem of clenbuterol toxic control is the lack of sensitive, specific and rapid methods for detection of clenbuterol residues.Monoclonal antibodies, which are of specific and identical, are concerned as optimum candidate in detection of clenbuterol residues.Clenbuterol is a hapten to prepare antigen,The clenbuterol was conjugated to carrier proteins KLH, BSA and OVA by diazotization. CL-KLH and CL-BSA were used for the immunization of mice and CL-OVA was used as the coating antigen for the ELISA study. Both antiserums of CL-KLH and CL-BSA could bind specifically with CL. The anti-clenbuterol antibody titer was higher in the sera of mice immunized with CL-KLH than that with CL-BSA.The Balb/c mice were immunized with CL-KLH. 4 strains of hybridoma cells, which secrete anti-CL antibody, named 1D6, 1H5, 1H7 and 2F10, were obtained by cell fusion and screening. The ascites of mAbs were prepared by injecting the hybridoma cells into mice abdomen. The indirect ELISA results showed that the McAbs revealed high affinity . The titers of McAb secreted by 4 strains of hybridorma are 1 : 3.2×105,l : 2.4×105,l : 4.8×104and 1 : 3.6×104,respectively. 1D6 has 3.98% cross reaction with Salbutamol. Relative affinity showed 1D6> 1H5> 1H7>2F1O.The competitive ELISA was developed to detect clenbuterol directly from the swine urine. Anti-CL antibody, named lD6,was employed to coat.treated sapmples added to the microplates. clenbuterol was diazotized to be joined with rabbit IgG. rabbit IgG-CL was used as the competitive...
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