Analytical and Bioanalytical Chemistry
Analytical and Bioanalytical Chemistry
4 years ago

A harmonized immunoassay with liquid chromatography-mass spectrometry analysis in egg allergen determination

Hideki Okada, Kazuhiko Kuroda, Tsutomu Honjoh, Kei Kurihara, Masaomi Nimata, Masahiro Shoji, Hirofumi Tachibana, Tsukasa Sugimoto, Takeo Yano

Abstract

Food allergy is a serious health issue worldwide. Implementing allergen labeling regulations is extremely challenging for regulators, food manufacturers, and analytical kit manufacturers. Here we have developed an “amino acid sequence immunoassay” approach to ELISA. The new ELISA comprises of a monoclonal antibody generated via an analyte specific peptide antigen and sodium lauryl sulfate/sulfite solution. This combination enables the antibody to access the epitope site in unfolded analyte protein. The newly developed ELISA recovered 87.1%–106.4% ovalbumin from ovalbumin-incurred model processed foods, thereby demonstrating its applicability as practical egg allergen determination. Furthermore, the comparison of LC-MS/MS and the new ELISA, which targets the amino acid sequence conforming to the LC-MS/MS detection peptide, showed a good agreement. Consequently the harmonization of two methods was demonstrated. The complementary use of the new ELISA and LC-MS analysis can offer a wide range of practical benefits in terms of easiness, cost, accuracy, and efficiency in food allergen analysis. In addition, the new assay is attractive in respect to its easy antigen preparation and predetermined specificity.

Graphical abstract

The ELISA composing of the monoclonal antibody targeting the amino acid sequence conformed to LC-MS detection peptide, and the protein conformation unfolding reagent was developed. In ovalbumin determination, the developed ELISA showed a good agreement with LC-MS analysis. Consequently the harmonization of immunoassay with LC-MS analysis by using common target amino acid sequence was demonstrated.

Publisher URL: https://link.springer.com/article/10.1007/s00216-017-0721-z

DOI: 10.1007/s00216-017-0721-z

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