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Enzyme-Linked Immunosorbent Assay01:33

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or...
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Identifying Protein-protein Interaction Sites Using Peptide Arrays
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Peptide array-based inhibition ELISA for evaluating antigenicity in infant formulas.

Chisato Kubo1, Masaki Kurimoto1, Masayoshi Tanaka2

  • 1Food Ingredients & Technology Institute, Morinaga Milk Industry Co., Ltd., 5-1-83 Higashihara, Zama, Kanagawa 252-8583, Japan.

Journal of Bioscience and Bioengineering
|July 28, 2020
PubMed
Summary

A new peptide array-based inhibition enzyme-linked immunosorbent assay (ELISA), Pep-iEIA, offers more sensitive detection of cow's milk allergens in infant formula. This method provides detailed analysis of remaining antigenic peptides, improving food safety for vulnerable populations.

Keywords:
Antigenicity evaluationInfant formulaInhibition enzyme-linked immunosorbent assayMilk allergyPeptide array

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Area of Science:

  • Food Science
  • Analytical Chemistry
  • Immunology

Background:

  • Consumer awareness of food safety and security necessitates robust food allergen control.
  • Existing methods for allergen detection often lack the required sensitivity for complex food matrices like infant formula.
  • Cow's milk allergy is a significant concern, particularly in infant populations, demanding accurate detection methods.

Purpose of the Study:

  • To develop and validate a highly sensitive assay for detecting cow's milk antigen in infant formula.
  • To evaluate the antigenicity of residual allergenic peptides in hydrolyzed infant formulas.
  • To provide a detailed analysis of allergenic peptide presence beyond conventional detection limits.

Main Methods:

  • Development of a peptide array-based inhibition enzyme-linked immunosorbent assay (Pep-iEIA) using overlapping peptides from allergenic milk proteins.
  • Application of Pep-iEIA to analyze five cow's milk-based infant formulas with varying degrees of hydrolysis.
  • Confirmation of identified allergenic peptide antigenicity using surface plasmon resonance (SPR) assay.

Main Results:

  • Pep-iEIA demonstrated superior sensitivity in detecting cow's milk antigen compared to conventional ELISA.
  • The assay provided a more detailed analysis of remaining antigenic peptides in hydrolyzed infant formulas.
  • Surface plasmon resonance confirmed the antigenicity levels of peptides identified by Pep-iEIA.

Conclusions:

  • Pep-iEIA is a highly sensitive and detailed method for evaluating antigenicity and detecting cow's milk allergens.
  • This technique is particularly valuable for assessing the safety of dairy products for infants and individuals with cow's milk allergy.
  • Pep-iEIA offers significant advancements in food allergen control and safety assurance in the food industry.