Abstract: Diethylstibestrol-4-Carboxymethyl ether was synthesized through chemical method in this paper, and was identified by nuclear magnetic resonance (NMR) and mass spectrometry (MS). To synthesize Diethylstibiestrol artificial antigen, Mixed anhydride and Isobutylchlorofomate and carbodiimide method were respectively used to conjugate Diethylstibestrol-4-Carboxymethyl ether to Bovine serum albumin (BSA) and Ovabumin (OVA). Diethylstibestrol artificial antigens were synthesized successfully, which were proved by ultraviolet scan、SDS-PAGE and animal experimentation. Anti-DES polyclonal antibody was prepared, and enzyme-linked immunosorbent assay method was founded. It’s very important to detect the residues of DES in food of animal origin.