| Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples |
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| 作者姓名： | 王文珺 凌云 许艇 高宏斌 生威 李季 |
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| 作者单位： | [1]College of Natural Resources and Environmental Science, China Agricultural University, Beijing 100094, China [2]College of Natural Sciences, China Agricultural University, Beijing 100094, China |
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| 基金项目： | Project supported by Ecology Key Subject of Beijing （No. XK10019440）. |
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| 摘 要： | 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-carboxypropyldiethylstilbestrol （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 carder 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 detection （LLD） of diethylstilbestrol was 0.01 ng/mL, and IC50= 1.02 ng/mL. Its analogs were tested and no obvious cross-reactivity was found to anti-diethylstilbestrol antibody. DES-fortified water samples were determined by simple dilution to diminish the matrix effect. The comparison between the amount of DES estimated by ELISA and the amount added indicates good agreement for all water samples tested, with mean recovery values ranging from 86% to 120.2%.
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| 关 键 词： | 乙烯雌酚 多克隆抗体 半抗原合成 ELISA 水样 基体干扰 |
| 修稿时间： | 2006-12-262007-03-27 |
Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples |
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| WANG Wen-Jun, LING Yun, XU Ting, GAO Hong-Bin,SHENG Wei,LI Ji.Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples[J].Chinese Journal of Chemistry,2007,25(8):1145-1150. |
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| Authors: | WANG Wen-Jun LING Yun XU Ting GAO Hong-Bin SHENG Wei LI Ji |
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| Institution: | 1. College of Natural Resources and Environmental Science, China Agricultural University, Beijing 100094, China ;2. College of Natural Sciences, China Agricultural University, Beijing 100094, China |
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| Abstract: | 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 detection (LLD) of diethylstilbestrol was 0.01 ng/mL, and IC50 = 1.02 ng/mL. Its analogs were tested and no obvious cross‐reactivity was found to anti‐diethylstilbestrol antibody. DES‐fortified water samples were determined by simple dilution to diminish the matrix effect. The comparison between the amount of DES estimated by ELISA and the amount added indicates good agreement for all water samples tested, with mean recovery values ranging from 86% to 120.2%. |
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| Keywords: | diethylstilbestrol polyclonal antibody hapten synthesis ELISA water sample matrix interference |
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