Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits

Cellerino Karina; Cagnasso Carolina Elisa; Greco Carola; Docena Guillermo; Polenta Gustavo; Ferreyra Verónica; López Laura Beatriz

doi:doi:10.11648/j.wjfst.20180201.12

Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits

Cellerino Karina, Cagnasso Carolina Elisa, Greco Carola, Docena Guillermo, Polenta Gustavo, Ferreyra Verónica, López Laura Beatriz

Published in World Journal of Food Science and Technology (Volume 2, Issue 1)

Received: 22 March 2018 Accepted: 8 April 2018 Published: 10 May 2018

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Abstract

Some protein ingredients declared in the label of gluten free products are allergenic proteins (milk, soy and egg). The proper identification of these proteins in food products is important for consumers who have food allergies. The aim of this study was to control the presence of protein ingredients declared in the label of gluten free products. Samples were analyzed with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), using an extractive solution of total proteins (Tris-ClH buffer 0.0625 M with 3% sodium dodecylsulfate and 2% 2-mercaptoethanol; pH: 6.8) and a selective solvent for the extraction of caseins (isopropanol 55° + 2-mercaptoethanol / ISO + ME). Developed Competitive enzyme immunoassays were used for the detection / quantification of milk, soy and egg proteins in products elaborated with rice flour. Specific polyclonal rabbit antiserums against milk, soy and egg proteins were used as primary antibodies in these competitive enzyme immunoassays. Commercial ELISA kits from Neogen, R-Biopharm and Romer were used to verify the results. In some samples undeclared allergens were detected. Correct allergens labeling is very important for the safety of allergic consumers. In conclusion, it is possible to identify all the proteins ingredients in these gluten-free foods studied, using a combination of electrophoretic methods and immunochemical methods.

Published in	World Journal of Food Science and Technology (Volume 2, Issue 1)
DOI	10.11648/j.wjfst.20180201.12
Page(s)	12-18
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), 2018. Published by Science Publishing Group

Keywords

Protein Ingredients, Allergenic Proteins, Gluten Free Products, SDS-PAGE, Developed Competitive Enzyme Immunoassays, Commercial ELISA Kit

References

[1]	Ward R. 2014. Chapter 1: Introduction to food aller-gy. In Flanagan S, Handbook of Food Allergen Detection and Control. Woodhead Publishing. Cambridge, UK.
[2]	Koeberl M., Clarke D., Allen K. J, Fleming F., Katzer L., Lee N. A., Lopata A. L., Said M., Scheelings p., Shepherd N., Sherlock R., Roberts J. 2018. Food Allergens Management in Australia. Journal of AOAC International. 101(1):60-69(10). DOI: https://doi.org/10.5740/jaoacint.17-0386
[3]	López M. C. 2018. Food Allergen Labeling: A Latin American Approach. Journal of AOAC International. 101(1):14-16(3). DOI: https://doi.org/10.5740/jaoacint.17-0382
[4]	Laemmli U. 1970. Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature. 227: 680-685.
[5]	López L. B. 2000. Separation, identification and quan-tification of proteins in processed foods. Thesis UBA. Argentina.
[6]	Cellerino K. 2016. Control Methodology for the analysis of milk, soy and egg allergens in meat and farinaceous products. Thesis UBA. Argentina.
[7]	Cellerino K., Rodríguez V., Docena G., López L. B. 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. Doi: 10.11648/j.jfns.20170502.16
[8]	R-Biopharm RIDASCREEN® FAST Milk. 2009. https://food.r-biopharm.com/wp-content/uploads/sites/2/2015/09/R4652-FAST-Milk-15-07-09.pdf. Access: january 2018.
[9]	Neogen Veratox® for Total milk allergen. 2016. http://foodsafety.neogen.com/pdf/procedures/8470_pro.pdf Access: january 2018.
[10]	R-Biopharm RIDASCREEN® FAST Soya. 2018. https://food.r-biopharm.com/wp-content/uploads/sites/2/2016/10/R7102-FAST-Soya-16-07-18.pdf. Access: january 2018.
[11]	Romer AgraQuant®. Soy Assay. 2011. http://shop.romerlabs.com/en/AgraQuant-ELISA/AgraQuant-Allergens/AgraQuant-ELISA-Soy. Access: january 2018.
[12]	Neogen Veratox® for Egg Allergen. 2013. http://foodsafety.neogen.com/en/veratox-egg. Access: january 2018.
[13]	Diaz Amigo C. 2010. Towards a Comprehensive Validation of ELISA Kits for Food Allergens. Case 2- Milk. Food Analytical Methods. 3: 351-356.
[14]	Do A. B, Khuda S. E, Sharma M. G. 2018. Undeclared food allergens and gluten in commercial food products analyzed by ELISA. Journal of AOAC International. 101(1):23-35(13). DOI: https://doi.org/10.5740/jaoacint.17-0384

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APA Style

Cellerino Karina, Cagnasso Carolina Elisa, Greco Carola, Docena Guillermo, Polenta Gustavo, et al. (2018). Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits. World Journal of Food Science and Technology, 2(1), 12-18. https://doi.org/10.11648/j.wjfst.20180201.12

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ACS Style

Cellerino Karina; Cagnasso Carolina Elisa; Greco Carola; Docena Guillermo; Polenta Gustavo, et al. Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits. World J. Food Sci. Technol. 2018, 2(1), 12-18. doi: 10.11648/j.wjfst.20180201.12

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AMA Style

Cellerino Karina, Cagnasso Carolina Elisa, Greco Carola, Docena Guillermo, Polenta Gustavo, et al. Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits. World J Food Sci Technol. 2018;2(1):12-18. doi: 10.11648/j.wjfst.20180201.12

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@article{10.11648/j.wjfst.20180201.12,
  author = {Cellerino Karina and Cagnasso Carolina Elisa and Greco Carola and Docena Guillermo and Polenta Gustavo and Ferreyra Verónica and López Laura Beatriz},
  title = {Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits},
  journal = {World Journal of Food Science and Technology},
  volume = {2},
  number = {1},
  pages = {12-18},
  doi = {10.11648/j.wjfst.20180201.12},
  url = {https://doi.org/10.11648/j.wjfst.20180201.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.wjfst.20180201.12},
  abstract = {Some protein ingredients declared in the label of gluten free products are allergenic proteins (milk, soy and egg). The proper identification of these proteins in food products is important for consumers who have food allergies. The aim of this study was to control the presence of protein ingredients declared in the label of gluten free products. Samples were analyzed with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), using an extractive solution of total proteins (Tris-ClH buffer 0.0625 M with 3% sodium dodecylsulfate and 2% 2-mercaptoethanol; pH: 6.8) and a selective solvent for the extraction of caseins (isopropanol 55° + 2-mercaptoethanol / ISO + ME). Developed Competitive enzyme immunoassays were used for the detection / quantification of milk, soy and egg proteins in products elaborated with rice flour. Specific polyclonal rabbit antiserums against milk, soy and egg proteins were used as primary antibodies in these competitive enzyme immunoassays. Commercial ELISA kits from Neogen, R-Biopharm and Romer were used to verify the results. In some samples undeclared allergens were detected. Correct allergens labeling is very important for the safety of allergic consumers. In conclusion, it is possible to identify all the proteins ingredients in these gluten-free foods studied, using a combination of electrophoretic methods and immunochemical methods.},
 year = {2018}
}

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TY  - JOUR
T1  - Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits
AU  - Cellerino Karina
AU  - Cagnasso Carolina Elisa
AU  - Greco Carola
AU  - Docena Guillermo
AU  - Polenta Gustavo
AU  - Ferreyra Verónica
AU  - López Laura Beatriz
Y1  - 2018/05/10
PY  - 2018
N1  - https://doi.org/10.11648/j.wjfst.20180201.12
DO  - 10.11648/j.wjfst.20180201.12
T2  - World Journal of Food Science and Technology
JF  - World Journal of Food Science and Technology
JO  - World Journal of Food Science and Technology
SP  - 12
EP  - 18
PB  - Science Publishing Group
SN  - 2637-6024
UR  - https://doi.org/10.11648/j.wjfst.20180201.12
AB  - Some protein ingredients declared in the label of gluten free products are allergenic proteins (milk, soy and egg). The proper identification of these proteins in food products is important for consumers who have food allergies. The aim of this study was to control the presence of protein ingredients declared in the label of gluten free products. Samples were analyzed with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), using an extractive solution of total proteins (Tris-ClH buffer 0.0625 M with 3% sodium dodecylsulfate and 2% 2-mercaptoethanol; pH: 6.8) and a selective solvent for the extraction of caseins (isopropanol 55° + 2-mercaptoethanol / ISO + ME). Developed Competitive enzyme immunoassays were used for the detection / quantification of milk, soy and egg proteins in products elaborated with rice flour. Specific polyclonal rabbit antiserums against milk, soy and egg proteins were used as primary antibodies in these competitive enzyme immunoassays. Commercial ELISA kits from Neogen, R-Biopharm and Romer were used to verify the results. In some samples undeclared allergens were detected. Correct allergens labeling is very important for the safety of allergic consumers. In conclusion, it is possible to identify all the proteins ingredients in these gluten-free foods studied, using a combination of electrophoretic methods and immunochemical methods.
VL  - 2
IS  - 1
ER  -

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Author Information

Cellerino Karina

Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina
Cagnasso Carolina Elisa

Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina
Greco Carola

Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina
Docena Guillermo

Institute of Immunologic and Physiopathologic Studies - IIFP, School of Sciences, National University of La Plata, Buenos Aires, Argentina
Polenta Gustavo

National Agricultural Technology Institute (INTA), Buenos Aires, Argentina
Ferreyra Verónica

Center for Research and Development in Industrialization Technology of Cereals and Oilseeds, National Institute of Industrial Technology, Buenos Aires, Argentina
López Laura Beatriz

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

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APA Style

Cellerino Karina, Cagnasso Carolina Elisa, Greco Carola, Docena Guillermo, Polenta Gustavo, et al. (2018). Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits. World Journal of Food Science and Technology, 2(1), 12-18. https://doi.org/10.11648/j.wjfst.20180201.12

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ACS Style

Cellerino Karina; Cagnasso Carolina Elisa; Greco Carola; Docena Guillermo; Polenta Gustavo, et al. Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits. World J. Food Sci. Technol. 2018, 2(1), 12-18. doi: 10.11648/j.wjfst.20180201.12

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AMA Style

Cellerino Karina, Cagnasso Carolina Elisa, Greco Carola, Docena Guillermo, Polenta Gustavo, et al. Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits. World J Food Sci Technol. 2018;2(1):12-18. doi: 10.11648/j.wjfst.20180201.12

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@article{10.11648/j.wjfst.20180201.12,
  author = {Cellerino Karina and Cagnasso Carolina Elisa and Greco Carola and Docena Guillermo and Polenta Gustavo and Ferreyra Verónica and López Laura Beatriz},
  title = {Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits},
  journal = {World Journal of Food Science and Technology},
  volume = {2},
  number = {1},
  pages = {12-18},
  doi = {10.11648/j.wjfst.20180201.12},
  url = {https://doi.org/10.11648/j.wjfst.20180201.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.wjfst.20180201.12},
  abstract = {Some protein ingredients declared in the label of gluten free products are allergenic proteins (milk, soy and egg). The proper identification of these proteins in food products is important for consumers who have food allergies. The aim of this study was to control the presence of protein ingredients declared in the label of gluten free products. Samples were analyzed with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), using an extractive solution of total proteins (Tris-ClH buffer 0.0625 M with 3% sodium dodecylsulfate and 2% 2-mercaptoethanol; pH: 6.8) and a selective solvent for the extraction of caseins (isopropanol 55° + 2-mercaptoethanol / ISO + ME). Developed Competitive enzyme immunoassays were used for the detection / quantification of milk, soy and egg proteins in products elaborated with rice flour. Specific polyclonal rabbit antiserums against milk, soy and egg proteins were used as primary antibodies in these competitive enzyme immunoassays. Commercial ELISA kits from Neogen, R-Biopharm and Romer were used to verify the results. In some samples undeclared allergens were detected. Correct allergens labeling is very important for the safety of allergic consumers. In conclusion, it is possible to identify all the proteins ingredients in these gluten-free foods studied, using a combination of electrophoretic methods and immunochemical methods.},
 year = {2018}
}

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TY  - JOUR
T1  - Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits
AU  - Cellerino Karina
AU  - Cagnasso Carolina Elisa
AU  - Greco Carola
AU  - Docena Guillermo
AU  - Polenta Gustavo
AU  - Ferreyra Verónica
AU  - López Laura Beatriz
Y1  - 2018/05/10
PY  - 2018
N1  - https://doi.org/10.11648/j.wjfst.20180201.12
DO  - 10.11648/j.wjfst.20180201.12
T2  - World Journal of Food Science and Technology
JF  - World Journal of Food Science and Technology
JO  - World Journal of Food Science and Technology
SP  - 12
EP  - 18
PB  - Science Publishing Group
SN  - 2637-6024
UR  - https://doi.org/10.11648/j.wjfst.20180201.12
AB  - Some protein ingredients declared in the label of gluten free products are allergenic proteins (milk, soy and egg). The proper identification of these proteins in food products is important for consumers who have food allergies. The aim of this study was to control the presence of protein ingredients declared in the label of gluten free products. Samples were analyzed with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), using an extractive solution of total proteins (Tris-ClH buffer 0.0625 M with 3% sodium dodecylsulfate and 2% 2-mercaptoethanol; pH: 6.8) and a selective solvent for the extraction of caseins (isopropanol 55° + 2-mercaptoethanol / ISO + ME). Developed Competitive enzyme immunoassays were used for the detection / quantification of milk, soy and egg proteins in products elaborated with rice flour. Specific polyclonal rabbit antiserums against milk, soy and egg proteins were used as primary antibodies in these competitive enzyme immunoassays. Commercial ELISA kits from Neogen, R-Biopharm and Romer were used to verify the results. In some samples undeclared allergens were detected. Correct allergens labeling is very important for the safety of allergic consumers. In conclusion, it is possible to identify all the proteins ingredients in these gluten-free foods studied, using a combination of electrophoretic methods and immunochemical methods.
VL  - 2
IS  - 1
ER  -

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