Development of an Indirect Competitive ELISA Based on.pdfVIP

Chinese Journal of Chemistry, 2007, 25, 1145— 1150 Full Paper Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples a b a WANG, Wen-Jun (王文珺) LING, Yun (凌云) XU, Ting (许艇) a a ,a GAO, Hong-Bin (高宏斌) SHENG, Wei (生威) LI, Ji* (李季) a College of Natural Resources and Environmental Science, China Agricultural University, Beijing 100094, China b College of Natural Sciences, China Agricultural University, Beijing 100094, China An indirect competitive enzyme-linked immunosorbent assay (icELISA) based on polyclonal antibody for the estrogen diethylstilbestrol (DES) was developed. With this aim, two different haptens mono-O-3-carboxypropyl- diethylstilbestrol (DES-CP) and mono-O-carboxymethyldiethylstilbestrol (DES-CM) with carboxylic group that preserve the molecular structure character of diethylstilbestrol were synthesized. The haptens were conjugated with the carrier proteins bovine serum albumin (BSA) by mixed-anhydride method for immunogen and conjugated with ovalbumin (OVA) by active ester method for coating antigen. Polyclonal antibodies for diethylstilbestrol were raised by immunizing mice with immune antigen DES-CP-BSA. Under optimized system, the lowest limit of detec-