| Clenbuterol Hydrochloride(CL), one of the β -adrenergic agonists, which is used to cure the bronchitis of human and livestock has a powerful effect of promoting animal growth. It was introduced as a growth-promoting agent to improve feed conversion efficiency and enhance lean meat to fat ration. However, accumulative residue of CL in food producing animals can cause symptoms of food poisoning in human.In this study, CL was linked with a new carrier(cationised bovine serum albumin) based on mannich reaction. 6-week-old female BALB/c mice were immunized intraperitoneally with this synthesized CL-cBSA. The spleen cells of immunized mice were fused with Sp2/0-Ag-14 myeloma cells. Indirect enzyme linked immunosorborbent assay (ELISA) was used to screen hybridoma cells and limited dilution method was performed to subclone positive clones.Two of McAbs specifically against clenbuterol were obtained and designed as 2E5 and 2H5. Both of them were IgM subtypes. The ELISA titers of ascitic fluids were 10 X2~9 and 10 X 2~11 respectively. The sensitivity and specificity was identified by a competitive inhibition enzyme-linked immunosorbent assay(ciELISA). The concentration range of CL from 102-10"2ng/ml, the inhibition rate of 2H5 to CL was higher than 2E5 obviously. The IC50 of 2H5 was lng/ml and 2E5 was more than 100ng/ml. The McAbs has no cross-reactivity towards Ractopamine and two carriers. The results showed that 2H5 was a good maretial to set up detecting method of CL.A competitive ELISA was established and detected the CL samples ranging fromlng/ml to 10ng/ml. 86 clinical urine samples were detected, among them 40 samples were positive and 46 samples were negative.
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