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Analytical and Bioanalytical Chemistry

, Volume 408, Issue 12, pp 3059–3069 | Cite as

Comprehensive peptide marker identification for the detection of multiple nut allergens using a non-targeted LC–HRMS multi-method

  • Robin Korte
  • Silke Lepski
  • Jens BrockmeyerEmail author
Paper in Forefront

Abstract

Food allergies have emerged as a global problem over the last few decades; therefore, reliable and sensitive analytical methods to ensure food safety for allergic consumers are required. The application of mass spectrometry is of growing interest in this field and several procedures based on low resolution tandem mass spectrometry using single tryptic peptides as analytical targets have recently been described. However, a comprehensive survey of marker peptides for the development of multi-methods is still missing, as is a consensus guide to marker identification. In this study, we therefore report a consistent approach to the development of liquid chromatography–mass spectrometry (LC–MS) multi-screening methods for the detection of allergens in food matrices. Proteotypic peptides were identified by a shotgun proteomics approach and verified through a thorough investigation of specificity and sensitivity. On the basis of this procedure, we identified 44 suitable tryptic marker peptides from six allergenic nut species and developed the first analytical LC–MS method for the detection of trace nut contaminations in processed foods using high resolution mass spectrometry (HRMS). The analysis of spiked matrix samples gave limits of detection (LODs) below 10 μg/g for several nuts; these LODs are comparable with routinely used methods such as ELISA and PCR. Notably, the HRMS approach can be used in an untargeted fashion to identify multiple allergens also retrospectively. In conclusion, we present here the so far largest consensus set of analytical markers from nut allergens and to the best of our knowledge the first multi-allergen method based on LC–HRMS.

Graphical Abstract

Identification of allergen peptide marker and LC–HRMS detection

Keywords

Food allergens High resolution mass spectrometry Orbitrap Multi-method Marker peptides Nut allergy 

Notes

Acknowledgments

The authors acknowledge support from the German National Academic Foundation (Studienstiftung des Deutschen Volkes) for funding this work and thank Katrin Bassen for providing the pictures for the graphical abstract.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

216_2016_9384_MOESM1_ESM.pdf (1.2 mb)
ESM 1 (PDF 1.24 mb)

References

  1. 1.
    Burks AW, Tang M, Sicherer S, Muraro A, Eigenmann PA, Ebisawa M, et al. ICON: food allergy. J Allergy Clin Immunol. 2012;129:906–20.CrossRefGoogle Scholar
  2. 2.
    Opara EI, Oehlschlager SL, Hanley AB. Immunoglobulin E mediated food allergy. Modelling and application of diagnostic and predictive tests for existing and novel foods. Biomarkers. 1998;3:1–19.CrossRefGoogle Scholar
  3. 3.
    Mills ENC, Breiteneder H. Food allergy and its relevance to industrial food proteins. Biotechnol Adv. 2005;23:409–14.CrossRefGoogle Scholar
  4. 4.
    Husain Z, Schwartz RA. Food allergy update: more than a peanut of a problem. Int J Dermatol. 2013;52:286–94.CrossRefGoogle Scholar
  5. 5.
    European Union (2011) Regulation (EU) No 1169/2011 of the European Parliament and of the Council of 25 October 2011 on the provision of food information to consumers. Off J Eur Union L 304/18:18–63Google Scholar
  6. 6.
    Costa J, Ansari P, Mafra I, Oliveira MBPP, Baumgartner S. Development of a sandwich ELISA-type system for the detection and quantification of hazelnut in model chocolates. Food Chem. 2015;173:257–65.CrossRefGoogle Scholar
  7. 7.
    Picariello G, Mamone G, Addeo F, Ferranti P. The frontiers of mass spectrometry-based techniques in food allergenomics. J Chromatogr A. 2011;1218:7386–98.CrossRefGoogle Scholar
  8. 8.
    Andjelkovic U, Martinovic T, Josic D. Foodomic investigations of food allergies. Curr Opin Food Sci. 2015;4:92–8.CrossRefGoogle Scholar
  9. 9.
    Chassaigne H, Nørgaard JV, Van Hengel AJ. Proteomics-based approach to detect and identify major allergens in processed peanuts by capillary LC-Q-TOF (MS/MS). J Agric Food Chem. 2007;55:4461–73.CrossRefGoogle Scholar
  10. 10.
    Walker MJ, Colwell P, Elahi S, Gray K, Lumley I. Food allergen detection: a literature review 2004–2007. J Assoc Public Anal. 2008;36:1–18.Google Scholar
  11. 11.
    Careri M, Costa A, Elviri L, Lagos JB, Mangia A, Terenghi M, et al. Use of specific peptide biomarkers for quantitative confirmation of hidden allergenic peanut proteins Ara h 2 and Ara h 3/4 for food control by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem. 2007;389:1901–7.CrossRefGoogle Scholar
  12. 12.
    Bignardi C, Elviri L, Penna A, Careri M, Mangia A. Particle-packed column versus silica-based monolithic column for liquid chromatography-electrospray-linear ion trap-tandem mass spectrometry multiallergen trace analysis in foods. J Chromatogr A. 2010;1217:7579–85.CrossRefGoogle Scholar
  13. 13.
    Heick J, Fischer M, Pöpping B. First screening method for the simultaneous detection of seven allergens by liquid chromatography mass spectrometry. J Chromatogr A. 2011;1218:938–43.CrossRefGoogle Scholar
  14. 14.
    Costa J, Ansari P, Mafra I, Oliveira MBPP, Baumgartner S. Assessing hazelnut allergens by protein- and DNA-based approaches: LC–MS/MS, ELISA and real-time PCR. Anal Bioanal Chem. 2014;406:2581–90.CrossRefGoogle Scholar
  15. 15.
    Monaci L, De Angelis E, Bavaro SL, Pilolli R. High-resolution Orbitrap™-based mass spectrometry for rapid detection of peanuts in nuts. Food Addit Contam Part A. 2015;32:1607–16.CrossRefGoogle Scholar
  16. 16.
    Chen Q, Zhang J, Ke X, Lai S, Tao B, Yang J, et al. Quantification of bovine β-casein allergen in baked foodstuffs based on ultra-performance liquid chromatography with tandem mass spectrometry. Food Addit Contam Part A. 2015;32:25–34.CrossRefGoogle Scholar
  17. 17.
    Monaci L, Losito I, Palmisano F, Godula M, Visconti A. Towards the quantification of residual milk allergens in caseinate-fined white wines using HPLC coupled with single-stage Orbitrap mass spectrometry. Food Addit Contam. 2011;28:1304–14.CrossRefGoogle Scholar
  18. 18.
    Monaci L, Losito I, De Angelis E, Pilolli R, Visconti A. Multi-allergen quantification of fining-related egg and milk proteins in white wines by high-resolution mass spectrometry. Rapid Commun Mass Spectrom. 2013;27:2009–18.CrossRefGoogle Scholar
  19. 19.
    Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.CrossRefGoogle Scholar
  20. 20.
    Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680–5.CrossRefGoogle Scholar
  21. 21.
    von Bargen C, Dojahn J, Waidelich D, Humpf H-U, Brockmeyer J. New sensitive high-performance liquid chromatography–tandem mass spectrometry method for the detection of horse and pork in halal beef. J Agric Food Chem. 2013;61:11986–94.CrossRefGoogle Scholar
  22. 22.
    Ma B, Zhang K, Hendrie C, Liang C, Li M, Doherty-Kirby A, et al. PEAKS: powerful software for peptide de novo sequencing by tandem mass spectrometry. Rapid Commun Mass Spectrom. 2003;17:2337–42.CrossRefGoogle Scholar
  23. 23.
    Zhang J, Xin L, Shan B, Chen W, Xie M, Yuen D, et al. PEAKS DB: de novo sequencing assisted database search for sensitive and accurate peptide identification. Mol Cell Proteomics. 2012;11:M111.010587.CrossRefGoogle Scholar
  24. 24.
    Makarov A, Denisov E, Lange O, Horning S. Dynamic range of mass accuracy in LTQ Orbitrap hybrid mass spectrometer. J Am Soc Mass Spectrom. 2006;17:977–82.CrossRefGoogle Scholar
  25. 25.
    von Bargen C, Brockmeyer J, Humpf H-U. Meat authentication: a new HPLC–MS/MS based method for the fast and sensitive detection of horse and pork in highly processed food. J Agric Food Chem. 2014;62:9428–35.CrossRefGoogle Scholar
  26. 26.
    Wu X, Beecher GR, Holden JM, Haytowitz DB, Gebhardt SE, Prior RL. Lipophilic and hydrophilic antioxidant capacities of common foods in the United States. J Agric Food Chem. 2004;52:4026–37.CrossRefGoogle Scholar
  27. 27.
    Sathe SK, Venkatachalam M, Sharma GM, Kshirsagar HH, Teuber SS, Roux KH. Solubilization and electrophoretic characterization of select edible nut seed proteins. J Agric Food Chem. 2009;57:7846–56.CrossRefGoogle Scholar
  28. 28.
    Willison LN, Zhang Q, Su M, Teuber SS, Sathe SK, Roux KH. Conformational epitope mapping of Pru du 6, a major allergen from almond nut. Mol Immunol. 2013;55:253–63.CrossRefGoogle Scholar
  29. 29.
    Willison LN, Tripathi P, Sharma G, Teuber SS, Sathe SK, Roux KH. Cloning, expression and patient IgE reactivity of recombinant Pru du 6, an 11S globulin from almond. Int Arch Allergy Immunol. 2011;156:267–81.CrossRefGoogle Scholar
  30. 30.
    Wang F, Robotham JM, Teuber SS, Sathe SK, Roux KH. Ana o 2, a major cashew (Anacardium occidentale L.) nut allergen of the legumin family. Int Arch Allergy Immunol. 2003;132:27–39.CrossRefGoogle Scholar
  31. 31.
    Robotham JM, Hoffman GG, Teuber SS, Beyer K, Sampson HA, Sathe SK, et al. Linear IgE-epitope mapping and comparative structural homology modeling of hazelnut and English walnut 11S globulins. Mol Immunol. 2009;46:2975–84.CrossRefGoogle Scholar
  32. 32.
    Ansari P, Stoppacher N, Baumgartner S. Marker peptide selection for the determination of hazelnut by LC–MS/MS and occurrence in other nuts. Anal Bioanal Chem. 2012;402:2607–15.CrossRefGoogle Scholar
  33. 33.
    Burks AW, Shin D, Cockrell G, Stanley JS, Helm RM, Bannon GA. Mapping and mutational analysis of the IgE-binding epitopes on Ara h 1, a legume vicilin protein and a major allergen in peanut hypersensitivity. Eur J Biochem. 1997;245:334–9.CrossRefGoogle Scholar
  34. 34.
    Shefcheck KJ, Callahan JH, Musser SM. Confirmation of peanut protein using peptide markers in dark chocolate using liquid chromatography-tandem mass spectrometry (LC–MS/MS). J Agric Food Chem. 2006;54:7953–9.CrossRefGoogle Scholar
  35. 35.
    Rabjohn P, Helm EM, Stanley JS, West CM, Sampson HA, Burks AW, et al. Molecular cloning and epitope analysis of the peanut allergen Ara h 3. J Clin Invest. 1999;103:535–42.CrossRefGoogle Scholar
  36. 36.
    Wang F, Robotham JM, Teuber SS, Tawde P, Sathe SK, Roux KH. Ana o 1, a cashew (Anacardium occidental) allergen of the vicilin seed storage protein family. J Allergy Clin Immunol. 2002;110:160–6.CrossRefGoogle Scholar
  37. 37.
    Ahn K, Bardina L, Grishina G, Beyer K, Sampson HA. Identification of two pistachio allergens, Pis v 1 and Pis v 2, belonging to the 2S albumin and 11S globulin family. Clin Exp Allergy. 2009;39:926–34.CrossRefGoogle Scholar
  38. 38.
    Jain AK (2004) Cloning and structural analysis of a cDNA clone encoding glycinin (Gly-1) seed storage protein of peanut. Electron J Biotechnol 7(3). doi: 10.2225/vol7-issue3-fulltext-13
  39. 39.
    Kang IH, Gallo M. Cloning and characterization of a novel peanut allergen Ara h 3 isoform displaying potentially decreased allergenicity. Plant Sci. 2007;172:345–53.CrossRefGoogle Scholar
  40. 40.
    Li HG, Wang L, Zhang YS, Lin XD, Liao B, Yan YS, et al. Cloning and sequencing of the gene Ahy-beta encoding a subunit of peanut conarachin. Plant Sci. 2005;168:1387–92.CrossRefGoogle Scholar
  41. 41.
    Barre A, Sordet C, Culerrier R, Rancé F, Didier A, Rougé P. Vicilin allergens of peanut and tree nuts (walnut, hazelnut and cashew nut) share structurally related IgE-binding epitopes. Mol Immunol. 2008;45:1231–40.CrossRefGoogle Scholar
  42. 42.
    Rougé P, Culerrier R, Sabatier V, Granier C, Rancé F, Barre A. Mapping and conformational analysis of IgE-binding epitopic regions on the molecular surface of the major Ara h 3 legumin allergen of peanut (Arachis hypogaea). Mol Immunol. 2009;46:1067–75.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Department of Chemistry, Institute of Food ChemistryWestfälische Wilhelms-Universität MünsterMünsterGermany

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