The Use of Combinatorial Hexapeptide Ligand Library (CPLL) in Allergomics

  • Youcef Shahali
  • Hélène Sénéchal
  • Pascal Poncet
Part of the Methods in Molecular Biology book series (MIMB, volume 1871)


The recent progress of proteomic protocols led to more efficient protein extraction and concentration procedures to remove nonprotein interfering compounds present in the starting material and to increase the concentration of underrepresented proteins. Combinatorial hexapeptide ligand libraries (CPLL) were recently applied to both plant- and animal-derived tissues for capturing the low- and very low-abundance allergens. Several IgE-binding proteins which were previously absent or poorly represented by using conventional proteomics tools have been detected and characterized through a CPLL-based approach. In the present chapter, a protocol based on improved protein extraction and enrichment by CPLL, allowing the immunochemical characterization of several “hidden allergens” in cypress pollen, is described in detail.

Key words

Hexapeptide ligand libraries Low-abundance allergens Pollen allergens Mass spectrometry Proteomics 



Collaboration with Drs. Egisto Boschetti and Pier Giorgio Righetti, the pioneer developers of the CPLL technology, is gratefully acknowledged and was at the basis of this protocol and its adaptation to allergomics studies.


  1. 1.
    Boschetti E, Righetti PG (2013) Low-abundance proteome discovery: state of the art and protocols. NewnesGoogle Scholar
  2. 2.
    Shahali Y, Sutra JP, Peltre G, Charpin D, Sénéchal H, Poncet P (2010) IgE reactivity to common cypress (C. sempervirens) pollen extracts: evidence for novel allergens. World Allergy Organ J 3:229–234CrossRefGoogle Scholar
  3. 3.
    Shahali Y, Sutra JP, Fasoli E, D’Amato A, Righetti PG, Futamura N et al (2012) Allergomic study of cypress pollen via combinatorial peptide ligand libraries. J Proteome 21:101–110CrossRefGoogle Scholar
  4. 4.
    Righetti PG, Fasoli E, D’Amato A, Boschetti E (2014) The “dark side” of food stuff proteomics: the CPLL-marshals investigate. Foods 3:217–237CrossRefGoogle Scholar
  5. 5.
    D’Amato A, Bachi A, Fasoli E, Boschetti E, Peltre G, Sénéchal H et al (2009) In-depth exploration of cow’s whey proteome via combinatorial peptide ligand libraries. J Proteome Res 8:3925–3936CrossRefGoogle Scholar
  6. 6.
    D'Amato A, Bachi A, Fasoli E, Boschetti E, Peltre G, Sénéchal H et al (2010) In-depth exploration of Hevea brasiliensis latex proteome and “hidden allergens” via combinatorial peptide ligand libraries. J Proteome 73:1368–1380CrossRefGoogle Scholar
  7. 7.
    Fasoli E, Pastorello EA, Farioli L, Scibilia J, Aldini G, Carini M et al (2009) Searching for allergens in maize kernels via proteomic tools. J Proteome 72:501–510CrossRefGoogle Scholar
  8. 8.
    Pedreschi R, Nørgaard J, Maquet A (2012) Current challenges in detecting food allergens by shotgun and targeted proteomic approaches: a case study on traces of peanut allergens in baked cookies. Nutrients 4:132–150CrossRefGoogle Scholar
  9. 9.
    D’Amato A, Kravchuk AV, Bachi A, Righetti PG (2010) Noah’s nectar: the proteome content of a glass of red wine. J Proteome 73:2370–2377CrossRefGoogle Scholar
  10. 10.
    Fekkar A, Pionneau C, Brossas JY, Marinach-Patrice C, Snounou G, Brock M, Mazier D (2012) DIGE enables the detection of a putative serum biomarker of fungal origin in a mouse model of invasive aspergillosis. J Proteome 75:2536–2549CrossRefGoogle Scholar
  11. 11.
    Martos G, López-Fandiño R, Molina E (2013) Immunoreactivity of hen egg allergens: influence on in vitro gastrointestinal digestion of the presence of other egg white proteins and of egg yolk. Food Chem 136:775–781CrossRefGoogle Scholar
  12. 12.
    Hartwig S, Lehr S (2012) Combination of highly efficient hexapeptide ligand library-based sample preparation with 2D DIGE for the analysis of the hidden human serum/plasma proteome. Methods Mol Biol 854:169–180CrossRefGoogle Scholar
  13. 13.
    Shahali Y, Nicaise P, Brazdova A, Charpin D, Scala E, Mari A et al (2014) Complementarity between microarray and immunoblot for the comparative evaluation of IgE repertoire of French and Italian cypress pollen allergic patients. Folia Biologica (Prague) 60:192Google Scholar
  14. 14.
    Danti R, Della Rocca G, Calamassi R, Mori B, Mariotti Lippi M (2011) Insights into a hydration regulating system in Cupressus pollen grains. Ann Bot 108:299–306CrossRefGoogle Scholar
  15. 15.
    Shahali Y (2011) Etude analytique de l’allergie au pollen de cyprès: aspects moléculaires et particulaires, Thesis Université Paris VI, Pierre et Marie Curie, Paris, France, p 220Google Scholar
  16. 16.
    Fasoli E, Farinazzo A, Sun CJ, Kravchuk AV, Guerrier L, Fortis F et al (2010) Interaction among proteins and peptide libraries in proteome analysis: pH involvement for a larger capture of species. J Proteome 73:733–742CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Youcef Shahali
    • 1
  • Hélène Sénéchal
    • 2
  • Pascal Poncet
    • 2
    • 3
  1. 1.Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension organization (AREEO)KarajIran
  2. 2.Armand Trousseau Children Hospital, AP-HP, Biochemistry Department, Allergy and Environment TeamParisFrance
  3. 3.Institut Pasteur, Center for Innovation and Technological ResearchParisFrance

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