Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples

Wang, Wenjun; Ling, Yun; Xu, Tao; Gao, Hongbin; Sheng, Wei; Li, Ji

doi:10.1002/cjoc.200790214

Cited by 17 publications

(8 citation statements)

References 29 publications

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“…Comparing the published ELISA methods (IC 50 = 1.02 μg L -1 [14]; IC 50 = 1.89 μg L -1 and LOD = 0.08 μg L -1 [15]), the antibody obtained by us demonstrated remarkably higher sensitivity. DES was able to compete with the coating antigen to interact with the antibody in the range of 50-1350 pg mL -1 , and in this range the absorbance value exhibited a good linear relationship with a logarithmic DES concentration (R 2 = 0.993; Figure 5).…”

Section: Antibody Evaluationmentioning

confidence: 87%

See 1 more Smart Citation

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Meng

He³

et al. 2011

Chin. Sci. Bull.

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Based on the preparation of an anti-diethylstilbestrol (DES) monoclonal antibody, a simple and convenient indirect competitive enzyme-linked immunosorbent assay (ELISA) method for DES detection has been developed. The monoclonal antibody demonstrated high sensitivity to DES with an IC 50 value of 275 pg mL -1 and detection limit (LOD) of 90 pg mL -1 . The specificity of the assay was studied by measuring cross-reactivity of the antibody with structurally related compounds of ethinyl estradiol (<7%), estrone (<0.1%), estriol (<0.1%), and diethylstilbestrol benzoate (<0.1%). Chicken, fish, shrimp, urine and bile spiked with different concentration of DES were detected by the developed method, and the recovery rates were greater than 79.5%. Intra-and inter-assay variations were about 6%. This method exhibited high stability with a coefficient of variation less than 10% in buffer and in real samples. The LODs in fish/shrimp, liver, feed and urine spiked with DES were 600, 600, 4800 and 600 pg mL -1 , respectively. These results confirmed that the antibody to DES was successfully produced and could be used to establish ELISA methods for DES detection in food producing animals. Diethylstilbestrol (DES; Figure 1) is an orally active synthetic non-steroid estrogen. Since the 1960s, DES has been used as a growth-promoting agent in livestock [1,2]. Later, DES was found to cause cancer and its use was "phased out in the 1970s" by the FAO/WHO. In 1971, DES was reported to be a teratogen when given to pregnant women and has been associated with a rare vaginal cancer in female offspring [3]. On February 5, 1975, the US Food and Drug Administration (FDA) ordered 25 and 100 mg tablets of DES withdrawn; however, it was still illegally used in the livestock industry.*Corresponding authors (email: xirimo2000@yahoo.com; jintliu@sdu.edu.cn)More than 30 years of researches have confirmed that DES is a teratogen, which can cause malformations of an embryo or fetus. Prenatal exposure to DES is associated with an increased incidence of fibroids of adulthood [4,5]. Therefore, the Ministry of Agriculture of the People's Republic of China issued a banned list (Announcement No. 235; http://www. agri.gov.cn/ zcfg/bmgz/ t20060123_540865.htm, accessed on October 24, 2002) of veterinary drugs and announced that the use of DES in food and animal feed was prohibited. The European Union (EU) requires that residual levels of DES in edible animal food should not exceed 2 ng mL -1 [6]. Consequently, a rapid screening method with high sensitivity and

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Section: Antibody Evaluationmentioning

confidence: 87%

“…Wang et al [14,15] produced two kinds of similar polyclonal antibodies against DES with IC 50 of 1.02 and 1.89 μg L -1 , respectively. Xu et al [16,17] also prepared a polyclonal antibody against DES with an average IC 50 value of 2.4 ng mL -1 , a calibration range of 0.2-30.5 ng mL -1 , and a detection limit of 0.07 ng mL -1 .…”

mentioning

confidence: 99%

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Meng

He³

et al. 2011

Chin. Sci. Bull.

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“…39 Hydrophilic molecularly imprinted polymers were prepared via bulk polymerization combined with hydrolysis of ester groups for selective recognition of iridoid glycosides. 40,41 In the work reported here, two types of MMIPs were synthesized by bulk polymerization and sol-gel methods, which used DES as the target template molecule, methacrylic acid (MAA) as a functional monomer and tetraethoxysiloxane (TEOS) as a crosslinking agent using a surface imprinting technique. The prepared MMIPs had excellent magnetic properties and good selectivity, and could specifically adsorb and rapidly separate the target from a complex food sample matrix under the action of a magnetic field.…”

Section: Introductionmentioning

confidence: 99%

Preparation of magnetic molecularly imprinted polymers for the rapid detection of diethylstilbestrol in milk samples

Huang

Zhao

2019

J Sci Food Agric

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BACKGROUND: Diethylstilbestrol (DES) residues are harmful to human health because of their potential carcinogenic properties. Therefore, it is important to develop a fast and efficient pretreatment method to prevent their harmful effects on human health and the environment. RESULTS: In this paper, two types of magnetic molecularly imprinted polymers (MMIPs) of DES were prepared by bulk polymerization and the sol-gel method, respectively. The synthetic materials were characterized using Fourier transform infrared spectroscopy, scanning electron microscopy and thermogravimetric analysis. Adsorption capacities of the bulk and sol-gel MMIPs were investigated. A rapid detection method was developed using the two types of MMIPs as sorbents, coupled to high-performance liquid chromatography, for the determination of DES residues in milk samples. Under optimized conditions, the limit of detection (S/N = 3) of both methods for DES was 2.0 g L −1 ; and the linear response range to DES was 0.1-500 mg L −1 . The milk samples were analyzed according to this method with good recoveries of 88.3-97.6 and 90.5-103.5% for the two types of MMIPs, respectively. CONCLUSIONS: The method described had high sensitivity and high selectivity, and could prove to be a new method for the rapid determination of DES residues in milk samples.where C 0 (μg mL −1 ) is the initial concentration of DES in acetonitrile solution, C (μg mL −1 ) is the equilibrium concentration of DES in acetonitrile solution, V (mL) is the volume of solution and W is the mass of MMIPs/MNIPs (g).J Sci Food Agric 2019; 99: 4452-4459Adsorption kinetics experiments Figure 6 shows the adsorption isotherms of MMIPs and MNIPs for DES prepared by bulk polymerization and the sol-gel method. The adsorption capacity of bulk and sol-gel MMIPs for DES increased J Sci Food Agric 2019; 99: 4452-4459

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“…[6][7][8][9][10][11] As a result, the use of DES in animal production was forbidden worldwide. 12 Various approaches for the determination of DES have been developed, such as immunoassay (ELISA), [13][14][15] gas chromatography-mass spectrometry (GC-MS) 16,17 and liquid chromatography-mass spectrometry (LC-MS). 18,19 However, ELLISA can be used only as a screening method because it tends to overestimate the concentration.…”

Section: Introductionmentioning

confidence: 99%

Determination of Diethylstilbestrol in Aquatic Products by High Performance Liquid Chromatography with Fluorescence Detection

Geng

Zhang

et al. 2010

Chin. J. Chem.

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A sensitive method for the determination of diethylstilbestrol (DES) in aquatic products by high performance liquid chromatography (HPLC) using a fluorescence detector (FLD) was developed. By means of sulfonation in concentrated sulfuric acid and hydrolysis, DES was converted into a reaction product that can emit fluorescence and determined by HPLC. For the first time, HPLC with FLD was applied to the determination of DES and sensitivity comparable to that of LC-MS gained. The limit of detection was 0.1 µg•kg -1 , and the recoveries were above 86%with relative standard deviations less than 6.2%. The proposed method was successfully applied to the determination of DES in aquatic products.

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Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples

Cited by 17 publications

References 29 publications

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Preparation of magnetic molecularly imprinted polymers for the rapid detection of diethylstilbestrol in milk samples

Determination of Diethylstilbestrol in Aquatic Products by High Performance Liquid Chromatography with Fluorescence Detection

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