Preparation of a monoclonal antibody and development of an indirect competitive ELISA for the detection of chlorpromazine residue in chicken and swine liver

Liu, Wei; Li, Weihua; Yin, Weiwei; Meng, Meng; Yue, Wan; Chen, Feng; Wang, Shanliang; Xi, Rimo

doi:10.1002/jsfa.4014

Cited by 8 publications

(17 citation statements)

References 22 publications

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“…Antiserum was collected and their titers were evaluated by an indirect competitive ELISA method. After the immune response was confirmed, the splenocytes from immunized mice were fused with mouse myeloma cells Sp2/0 (7 : 1, cell ratio) using the polyethylene glycol (PEG) method [18] and cells were suspended in hypoxanthineaminopterin-thymidine medium. The hybridoma cells were cultured in medium (pH 7.4) containing 0.2% NaHCO 3 and RPMI1640 with the addition of 20% newborn calf serum at 37°C to obtain MAbs for DES.…”

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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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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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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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“…detecting the pathogens at low detection limit (Liu et al, 2010). In contrast, nowadays, the demand for immunoglubulin (IgY) is increasing not only for immunological purposes but also for therapeutic applications (Carlander et al, 2003).…”

Section: Articlementioning

confidence: 99%

Development of Chicken Immunoglobulin Y for Rapid Detection of Cronobacter muytjensii in Infant Formula Powder

Kim¹,

Shukla̽²,

Ahmed³

et al. 2012

Korean Journal for Food Science of Animal Resources

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The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 µg/mL, having ca. 10 8 coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of 10 5 CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

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“…Immunoassays are a class of low cost, rapid, and sensitive methods capable of screening relatively large numbers of samples in a single test. By now, there have been some immunoassays reported for the determination of PZs, [15][16][17][18][19][20][21][22] but all these methods have their respective disadvantages.…”

Section: Introductionmentioning

confidence: 99%

Synthesis of novel hapten and production of the generic monoclonal antibody for phenothiazine drugs

Liu

Gao

Zhang

et al. 2014

Journal of Environmental Science and Health, Part B

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The objective of the present study was to produce a generic monoclonal antibody for phenothiazine drugs. Phenothiazine was derivatized with sodium chloroacetate to synthesize a generic hapten that was then coupled with bovine serum albumin to prepare the immunogen by use of n-hydroxysuccinimide active ester method. Hybridomas were obtained by fusing mouse myeloma cells SP2/0 with the splenocyte cells from the mice immunized with the immunogen. Then, a competitive indirect enzyme-linked immunosorbent assay was used to determine the recognization capacity of the obtained monoclonal antibody for phenothiazine drugs. Results showed that the monoclonal antibody of phenothiazine simultaneously recognized five phenothiazine drugs (chlorpromazine, promethazine, acepromazin, perphenazine, and fluphenazine). The cross-reactivities were in the range of 42%-87% and the limits of detection were in the range of 0.2-0.7 ng/mL. Therefore, the obtained monoclonal antibody in this study was suitable for the development of an immunoassay to multi-detect the five phenothiazine drugs.

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Preparation of a monoclonal antibody and development of an indirect competitive ELISA for the detection of chlorpromazine residue in chicken and swine liver

Cited by 8 publications

References 22 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

Development of Chicken Immunoglobulin Y for Rapid Detection of Cronobacter muytjensii in Infant Formula Powder

Synthesis of novel hapten and production of the generic monoclonal antibody for phenothiazine drugs

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