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Proteomic Analysisof Food Allergens by MALDI TOF/TOF Mass Spectrometry

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Protein Downstream Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2178))

Abstract

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is largely recognized as an important tool in the analysis of many biomolecules such as proteins and peptides. The MS analysis of digested peptides to identify a protein or some of its modifications is a key step in proteomics. MALDI-MS is well suited for the peptide mass fingerprinting (PMF) technique, as well as selected fragmentation of various precursors using collisional-induced dissociation (CID) or post-source decay (PSD).

In the last few years, MALDI-MS has played a significant role in food chemistry, especially in the detection of food adulterations, characterization of food allergens, and investigation of protein structural modifications induced by various industrial processes that could be an issue in terms of food quality and safety.

Here, we present simple extraction protocols of allergenic proteins in food commodities such as milk, egg, hazelnut , and lupin seeds. Classic bottom-up approaches based on Sodium Dodecyl Sulphate (SDS) gel electrophoresis separation followed by in-gel digestion or direct in-solution digestion of whole samples are described. MALDI-MS and MS /MS analyses are discussed along with a comparison of data obtained by using the most widespread matrices for proteomic studies, namely, α-cyano-4-hydroxy-cinnamic acid (CHCA) and α-cyano-4-chloro-cinnamic acid (CClCA). The choice of the most suitable MALDI matrix is fundamental for high-throughput screening of putative food allergens.

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Acknowledgments

This work was supported by Progetto di Ricerca di Interesse Nazionale—PRIN 2017YER72K—“Development of novel DNA-based analytical platforms for the rapid, point-of-use quantification of multiple hidden allergens in food samples.”

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Authors and Affiliations

  1. Centro Interdipartimentale di Ricerca SMART, Università degli Studi di Bari “Aldo Moro”, Bari, Italy

    Cosima D. Calvano, Ilario Losito & Tommaso R. I. Cataldi

  2. Dipartimento di Farmacia- Scienze del Farmaco, Università degli Studi di Bari “Aldo Moro”, Bari, Italy

    Cosima D. Calvano

  3. Dipartimento di Chimica, Università degli Studi di Bari “Aldo Moro”, Bari, Italy

    Mariachiara Bianco, Ilario Losito & Tommaso R. I. Cataldi

Authors
  1. Cosima D. Calvano
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  2. Mariachiara Bianco
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  3. Ilario Losito
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  4. Tommaso R. I. Cataldi
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Corresponding author

Correspondence to Cosima D. Calvano .

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Editors and Affiliations

  1. Laboratory of Enzyme Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, Athens, Greece

    Nikolaos E. Labrou

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Calvano, C.D., Bianco, M., Losito, I., Cataldi, T.R.I. (2021). Proteomic Analysisof Food Allergens by MALDI TOF/TOF Mass Spectrometry. In: Labrou, N.E. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 2178. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0775-6_24

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  • DOI: https://doi.org/10.1007/978-1-0716-0775-6_24

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