| Diethylstilbestrol-mono-carboxylic propyl ether(DES-MCPE) was synthesized and purified.The haptens were covalently conjugated to carrier proteins BSA and OVA by the method of active ester to prepare immunogen and coating antigen.The conjugates were confirmed by UV spectrophotometry and SDA-PAGE.their molar ratios of haptens to carrier protein were 17.5:1 and 21:1.After immunization of Balb/c mice with synthesized artificial antigen diethylstilbestrol-bovine serum albumin(DES-BSA),the stimulated splenocytes were fused with SP2/0 myeloma cells to produce hybridomas,two hybridoma cell lines producing monoclonal antibody against diethylstilbestrol(DES)were established,Which was designated 1F5 and 2D8.The mice(BALB/c) were injected intraperitoneally with 1F5 and 2D8 hybridomas to produce ascites.the ascites was purified by(NH4)2SO4.The indirect ELISA titers of cell supernatant and ascites were 1:640,1:320and 3.2×106,1.6×106 respectively.The monoclonal antibodies were of subclasses IgG2a,The number of chromosome of the hybridoma cell lines was 99.24±2.5 and 99.45±1.5.The competitive inhibition enzyme-linked immunosorbent assay(ciELISA) for DES is eatablished with the ascites.the most suitable working condition of ciELISA is obtained by several grid tests which are used to calculate the most optimal concentration and dilution of the coating antigen and ascites,and the result is that the coating antigen concentration is 1μg/mL and the dilution of ascites is 1:3.2*105. After optimized some parameters such as the coating time,blocking solution, blocking time,incubation time,pH,obvious time and so on.finally,the regression equation of this curve is y=-25.629LogC+79.487,R2=0.9881.the curve has a favorable linearity relation within the concentration range of 1～100 ng/ml.The very high specificity of the McAb for DES was expressed by the insignificant cross-reactivity with other compounds and drugs.The average rate of recovery of muscle by ciELISA is108.6%,the result show that the accuracy is well.
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