Preparation and validation of the polyclonal antibodies for detection of chlortetracycline residues

Zhao, Chengbiao; Peng, Dengfeng; Wang, Y.L.; Huang, Lingli; Chen, D.M.; Tao, Yanfei; Zhang, Yuan

doi:10.1080/09540100802117875

Cited by 10 publications

(10 citation statements)

References 20 publications

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“…Most of the reported antibodies specific for TC group antibiotics are PAbs (Zhao et al, 2008;Le et al, 2009;Cháfer-Pericása et al, 2010). Because the properties of PAbs vary among batches according to the immune responses of different immunized animals, PAbs are less suitable for long-term application than MAbs (Hock et al, 1995).…”

Section: Production Of Mabsmentioning

confidence: 99%

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Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp

Wongtangprasert

Natakuathung

Pimpitak

et al. 2014

J. Zhejiang Univ. Sci. B

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A novel monoclonal antibody (MAb) against oxytetracycline (OTC) was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay (ELISA)-based detection system. An OTC-bovine serum albumin (BSA) conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2-4F, 7-3G, and 11-11A. An indirect competitive ELISA (icELISA) was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration (IC 50 ) and percentage of cross-reactivity, respectively. MAb 2-4F exhibited the highest sensitivity, with an IC 50 of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no crossreactivity to other unrelated antibiotics. When MAb 2-4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%-118% for an intra-assay and 96%-113% for an inter-assay. The coefficients of variation of the assays were 3.9%-13.9% and 5.5%-14.9%, respectively.

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Section: Production Of Mabsmentioning

confidence: 99%

“…It has been reported that sample matrixes could have an effect on the efficiency of an ELISA (Zhao et al, 2008). Therefore, sample preparation must be modified for other types of matrix.…”

Section: Otc Analysis In Fortified Shrimp Samplesmentioning

confidence: 99%

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp

Wongtangprasert

Natakuathung

Pimpitak

et al. 2014

J. Zhejiang Univ. Sci. B

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“…The limit of detection (90% of B/Bo) was 0.05 ng ml À1 (Figure 2). The entire protocol of the conventional indirect competitive ELISA developed by Zhao et al (2008) took at least 4 h, including three washing and three incubation steps, with IC 50 ¼ 15 ng ml À1 . In the present study the modified simplified ELISA only took 1 h with two washings and two incubations with an IC 50 of 0.66 ng ml À1 .…”

Section: Resultsmentioning

confidence: 99%

“…Immunogen (chlortetracycline-bovine serum albumin, CTC-BSA) and coating antigen (chlortetracyclineovalbumin CTC-OVA) were synthesized by EDC Coupling Method according to the procedure described previously with some modifications (Zhao et al 2008). A total of 10 -3 M CTC was dissolved in 30 ml PBS (0.01 mM, pH 7.4), then 10 À3 M sodium chloroacetate was added slowly into the solution.…”

Section: Immunogens and Coating Antigens Preparationmentioning

confidence: 99%

“…ELISA has proved to be a simple, rapid and costeffective method for monitoring trace residues of antibiotics, particularly for high-throughput and onsite screening analysis. Two ELISA methods based on polyclonal antibody were developed for the detection of CTC residues in animal tissues, but they have high cross-reactivity with other TCs and the detection limits were unsatisfactory (Everest et al 1994;Zhao et al 2008). Subsequently, three improved ELISA methods were developed by Pastor-Navarro et al (2007), Le et al (2009) and Zhang et al (2007) to detect TCs in honey or milk with 10% cross-reactivity to CTC or a high concentration at 50% inhibition (IC 50 ).…”

Section: Introductionmentioning

confidence: 99%

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Rapid and sensitive enzyme-linked immunosorbent assay and immunochromatographic assay for the detection of chlortetracycline residues in edible animal tissues

Zhao

et al. 2011

Food Additives & Contaminants: Part A

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Two rapid and sensitive enzyme-linked immunosorbent assays (ELISA) and an immunochromatographic assay (ICA) for the detection of chlortetracycline (CTC) residues in edible animal tissues were developed based on a monoclonal antibody (MAb) produced by using the chlortetracycline-bovine serum albumin (CTC-BSA) conjugate as the immunogen. A total of 50% inhibiting concentration (IC(50)) of the modified ELISA was 0.66 ng ml(-1) and the recoveries from spiked chicken muscle and liver were 78.8-92.2% and 80.3-90.2%, respectively. The corresponding coefficient variations (CVs) were 3.2-9.5% and 6.5-10.2%. The detection limit was 0.06 ng g(-1) in chicken muscle and 0.07 ng g(-1) in liver. However, the detection limit of ICA was 0.12 ng ml(-1), and the recoveries in negative samples spiked at concentrations of 10, 50 and 100 ng g(-1) ranged from 79.0% to 88.6% for muscle samples and from 75.2% to 87.0% for liver samples. The cut-off values for the test lines were 80 ng g(-1) and the analysis can be completed within 5-10 min. Comparisons with an HPLC method were performed by testing 200 swine muscle samples and chicken muscle samples from local markets, and an agreement rate of 99.5% was obtained between the three methods.

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A receptor-based chemiluminescence enzyme linked immunosorbent assay for determination of tetracyclines in milk

Wang

Zhang

Liu

et al. 2019

Analytical Biochemistry

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Preparation and validation of the polyclonal antibodies for detection of chlortetracycline residues

Cited by 10 publications

References 20 publications

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp

Rapid and sensitive enzyme-linked immunosorbent assay and immunochromatographic assay for the detection of chlortetracycline residues in edible animal tissues

A receptor-based chemiluminescence enzyme linked immunosorbent assay for determination of tetracyclines in milk

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