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Development of a Competitive Enzyme Immunoassay Technique for the Detection of Soy Traces in Meat Products

Received: 30 January 2017     Accepted: 22 March 2017     Published: 3 April 2017
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Abstract

The aim of this work was to develop a competitive enzyme immunoassay technique, to detect the presence of traces of soy in meat products. Specific rabbit polyclonal antiserum against soy protein was used as primary antibody. The optimal antigen concentration to be immobilized on the plate and the concentration of primary antibody to be used in competition was determined. The calibration curve was fitted using increasing concentrations of an extract of soy product. The soy product was extracted with Tris-HCl buffer 0.0625M with 3% sodium dodecylsulfate and 2% mercaptoethanol. The working range used in the enzyme immunoassay to detect soy was 9-280ppm SP with adequate linearity (R2: 0.9880). All validation parameters studied were appropriate. Commercial samples of meat products were analyzed with this enzyme immunoassays and a commercial ELISA kit. Significant differences were observed in the quantitative results obtained with both methods; nevertheless the developed enzyme immunoassay could be used as screening method.

Published in Journal of Food and Nutrition Sciences (Volume 5, Issue 2)
DOI 10.11648/j.jfns.20170502.16
Page(s) 57-62
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2017. Published by Science Publishing Group

Keywords

ELISA, Allergens, Soy Detection, Meat Products

References
[1] López, L. B., Greco, C. B., Ronayne de Ferrer, P., Valencia, M. E. 2006. Identification of extrinsic proteins in boneless cooked ham by SDS-PAGE: detection level in model systems. Archivos Latinoamericanos de Nutrición. 56 (3): 282.
[2] López L, Binaghi M, Greco C, Mambrín M. Cellerino K, Valencia M. 2010. Salted sausages and dry sausages: detection by electrophoresis of meat species and extrinsic aggregate proteins. DIAETA. 28 (131): 7-13.
[3] Ward R. 2014. Chapter 1: Introduction to food allergy. In Flanagan S, Handbook of Food Allergen Detection and Control. Woodhead Publishing. Cambridge, UK.
[4] Lowry O, Rosebrough N, Farr A, Randall R. 1951. Protein measurement with the Folin Phenol reagent. J Biol Chem. 193: 265–275.
[5] Olivera Carrión M. 1988. Separation of dietary proteins by electrophoresis: Study of the changes induced by the processing, identification of species and detection of proteins in mixtures. Thesis. UBA. Argentina.
[6] Rozenfeld P, Docena G, Añón M, Fossati C. 2002. Detection and identification of a SP component that cross reacts with caseins from cow milk. Clin Exp Immunol. 130 (1): 49-58.
[7] Box G, Hunter W, Stuart Hunter J. 1999. Statistics for researchers. Introduction to the design of experiments, data analysis and model construction. Editorial Reverté S. A., Mexico D. F.
[8] Huber L. 2010. Validation of Analytical Methods. Agilent Technologies, Germany.
[9] Gatti M y Ferretti C. 2010. Chapter 17: Soy Allergen Detection, en Popping B, Diaz Amigo C, Hoenicke K, Molecular Biological and immunological techniques and applications for food chemists. John Wiley & Sons, Inc., Canada.
[10] Ito K, Yamamoto T, Oyama Y, Tsuruma R, Saito E, Saito Y, Ozu T1, Honjoh T, Adachi R, Sakai S, Akiyama H, Shoji M. 2016. Food allergen analysis for processed food using a novel extraction method to eliminate harmful reagents for both ELISA and lateral-flow tests. Anal Bioanal Chem. 408 (22): 5973-84.
[11] Watanabe Y, Aburatani K, Mizumurz T, Sakai M, Muraoka S, Mamegosi S, Honjoh T. 2005. Novel ELISA for the detection of raw and processed egg using extraction buffer containing a surfactant and reducing agent. J Immunol Methods. 300: 115-123.
[12] Diaz Amigo C y Popping B. 2010. Analytical Testing as a Tool for the Enforcement of future Regulatory Thresholds for Food Allergens. Journal of AOAC International. 93 (2): 434-441.
[13] Cellerino Karina, Lopez Laura Beatriz, SP Detection in Raw and Cooked Meat Products Using Different ELISA Kits, Journal of Food and Nutrition Sciences. Vol. 4, No. 6, 2016, pp. 170-174. doi: 10.11648/j.jfns.20160406.15.
[14] Diaz Amigo C. 2010a. Capítulo 12: Antibody-Based Detection Methods: From Theory to Practice, en Popping B, Diaz-Amigo C, Hoenicke K, Molecular Biological and immunological techniques and applications for food chemists. John Wiley & Sons, Inc., Canada.
[15] Diaz Amigo C. 2010b. Towards a Comprehensive Validation of ELISA Kits for Food Allergens. Case 2- Milk. Food Analytical Methods. 3: 351-356.
Cite This Article
  • APA Style

    Cellerino Karina, Rodríguez Viviana Gladys, Docena Guillermo, López Laura Beatriz. (2017). Development of a Competitive Enzyme Immunoassay Technique for the Detection of Soy Traces in Meat Products. Journal of Food and Nutrition Sciences, 5(2), 57-62. https://doi.org/10.11648/j.jfns.20170502.16

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    ACS Style

    Cellerino Karina; Rodríguez Viviana Gladys; Docena Guillermo; López Laura Beatriz. Development of a Competitive Enzyme Immunoassay Technique for the Detection of Soy Traces in Meat Products. J. Food Nutr. Sci. 2017, 5(2), 57-62. doi: 10.11648/j.jfns.20170502.16

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    AMA Style

    Cellerino Karina, Rodríguez Viviana Gladys, Docena Guillermo, López Laura Beatriz. Development of a Competitive Enzyme Immunoassay Technique for the Detection of Soy Traces in Meat Products. J Food Nutr Sci. 2017;5(2):57-62. doi: 10.11648/j.jfns.20170502.16

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  • @article{10.11648/j.jfns.20170502.16,
      author = {Cellerino Karina and Rodríguez Viviana Gladys and Docena Guillermo and López Laura Beatriz},
      title = {Development of a Competitive Enzyme Immunoassay Technique for the Detection of Soy Traces in Meat Products},
      journal = {Journal of Food and Nutrition Sciences},
      volume = {5},
      number = {2},
      pages = {57-62},
      doi = {10.11648/j.jfns.20170502.16},
      url = {https://doi.org/10.11648/j.jfns.20170502.16},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jfns.20170502.16},
      abstract = {The aim of this work was to develop a competitive enzyme immunoassay technique, to detect the presence of traces of soy in meat products. Specific rabbit polyclonal antiserum against soy protein was used as primary antibody. The optimal antigen concentration to be immobilized on the plate and the concentration of primary antibody to be used in competition was determined. The calibration curve was fitted using increasing concentrations of an extract of soy product. The soy product was extracted with Tris-HCl buffer 0.0625M with 3% sodium dodecylsulfate and 2% mercaptoethanol. The working range used in the enzyme immunoassay to detect soy was 9-280ppm SP with adequate linearity (R2: 0.9880). All validation parameters studied were appropriate. Commercial samples of meat products were analyzed with this enzyme immunoassays and a commercial ELISA kit. Significant differences were observed in the quantitative results obtained with both methods; nevertheless the developed enzyme immunoassay could be used as screening method.},
     year = {2017}
    }
    

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    T1  - Development of a Competitive Enzyme Immunoassay Technique for the Detection of Soy Traces in Meat Products
    AU  - Cellerino Karina
    AU  - Rodríguez Viviana Gladys
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    AU  - López Laura Beatriz
    Y1  - 2017/04/03
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    DO  - 10.11648/j.jfns.20170502.16
    T2  - Journal of Food and Nutrition Sciences
    JF  - Journal of Food and Nutrition Sciences
    JO  - Journal of Food and Nutrition Sciences
    SP  - 57
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    SN  - 2330-7293
    UR  - https://doi.org/10.11648/j.jfns.20170502.16
    AB  - The aim of this work was to develop a competitive enzyme immunoassay technique, to detect the presence of traces of soy in meat products. Specific rabbit polyclonal antiserum against soy protein was used as primary antibody. The optimal antigen concentration to be immobilized on the plate and the concentration of primary antibody to be used in competition was determined. The calibration curve was fitted using increasing concentrations of an extract of soy product. The soy product was extracted with Tris-HCl buffer 0.0625M with 3% sodium dodecylsulfate and 2% mercaptoethanol. The working range used in the enzyme immunoassay to detect soy was 9-280ppm SP with adequate linearity (R2: 0.9880). All validation parameters studied were appropriate. Commercial samples of meat products were analyzed with this enzyme immunoassays and a commercial ELISA kit. Significant differences were observed in the quantitative results obtained with both methods; nevertheless the developed enzyme immunoassay could be used as screening method.
    VL  - 5
    IS  - 2
    ER  - 

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Author Information
  • Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina

  • Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina

  • Institute of Immunologic and Physiopathologic Studies - IIFP, School of Sciences, National University of La Plata, UNLP, La Plata, Argentina

  • Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina

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