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Preparation of Artificial Antigen and Development of Indirect Competitive ELISA Based on Chicken IgY for the Detection of Acid Orange II in Food Samples

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Abstract

To evaluate the feasibility of specific egg yolk antibodies (IgY) technology for the detection of hazardous chemical residues in foodstuffs, IgY were generated to detect acid orange II (AO2) residues. Bovine serum albumin (BSA) was made to bind with AO2 by succinic anhydride (SA), and the conjugate was used to immunize the laying chickens. PEG-6000 precipitation was used to extract IgY antibodies. The titer of anti-AO2 IgY attained the peak (1:12,800) after fifth booster immunization. Checkerboard titration showed that 1:640 dilution of anti-AO2 IgY could approximately give an optical density (OD) 1.0 at 5 μg/mL AO2-OVA coating concentration. The anti-AO2 IgY based indirect competitive ELISA (ic-ELISA) showed that the IC50 value of anti-AO2 IgY was 10.72 μg/mL and regression curve equation was y =−24.41 × +75.11 (R 2 = 0.946). The ic-ELISA showed less cross-reactivity in range between 0.09 and 21.07 %. Recoveries from dried bean curd, sausage, and chilli powder samples were from 78.80 to 152.90 %, with relative standard deviation lower than 19.13 %. The study suggests that generated anti-AO2 IgY antibodies could be used in routine screening analysis of AO2 residues in food samples.

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Acknowledgements

This work was supported by the Ph.D. Programs Foundation of Ministry of Education of China (20130204110023); Key Construction Program of International Cooperation Base in S&T, (2015SD0018) Shaanxi Province, China; Ministry of Education and State Administration of Foreign Experts Affairs “Overseas Teacher” project (No.MS2011XBNL057), China.

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Conflict of Interest

Jinxin He declares that he has no conflict of interest.

Yuan Wang declares that he has no conflict of interest.

Xiaoying Zhang declares that he has no conflict of interest.

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

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Authors and Affiliations

  1. College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, People’s Republic of China

    Jinxin He, Yuan Wang & Xiaoying Zhang

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  1. Jinxin He
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  2. Yuan Wang
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  3. Xiaoying Zhang
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Correspondence to Xiaoying Zhang.

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He, J., Wang, Y. & Zhang, X. Preparation of Artificial Antigen and Development of Indirect Competitive ELISA Based on Chicken IgY for the Detection of Acid Orange II in Food Samples. Food Anal. Methods 9, 378–384 (2016). https://doi.org/10.1007/s12161-015-0203-x

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  • DOI: https://doi.org/10.1007/s12161-015-0203-x

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