Analysis of Calcium-Induced Conformational Changes in Calcium-Binding Allergens and Quantitative Determination of Their IgE Binding Properties

  • Nuria Parody
  • Miguel Ángel Fuertes
  • Carlos Alonso
  • Yago Pico de Coaña
Part of the Methods in Molecular Biology book series (MIMB, volume 963)


The polcalcin family is one of the most epidemiologically relevant families of calcium-binding allergens. Polcalcins are potent plant allergens that contain one or several EF-hand motifs and their allergenicity is primarily associated with the Ca2+-bound form of the protein. Conformation, stability, as well as IgE recognition of calcium-binding allergens greatly depend on the presence of protein-bound calcium ions.

We describe a protocol that uses three techniques (SDS-PAGE, circular dichroism spectroscopy, and ELISA) to describe the effects that calcium has on the structural changes in an allergen and its IgE binding properties.

Key words

Allergen Polcalcin Calcium EF-hand IgE binding 


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Nuria Parody
    • 1
    • 2
  • Miguel Ángel Fuertes
    • 1
  • Carlos Alonso
    • 1
  • Yago Pico de Coaña
    • 1
  1. 1.Centro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones CientificasUniversidad Autonoma de MadridMadridSpain
  2. 2.R&D DepartmentLaboratorios LETI S.LMadridSpain

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