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Molecular Biotechnology

, Volume 56, Issue 6, pp 498–506 | Cite as

Molecular Characterization of Recombinant Mus a 5 Allergen from Banana Fruit

  • Ivan Mrkic
  • Mohamed Abughren
  • Jasna Nikolic
  • Uros Andjelkovic
  • Emilia Vassilopoulou
  • Athanassios Sinaniotis
  • Arnd Petersen
  • Nikolaos G. Papadopoulos
  • Marija Gavrovic-JankulovicEmail author
Research

Abstract

Allergy to banana fruit appears to have become an important cause of fruit allergy in Europe. Among five allergens that have been found, beta-1,3-glucanase denoted as Mus a 5 was identified as a candidate allergen for the component-resolved allergy diagnosis of banana allergy. Because of the variations in protein levels in banana fruit, in this study Mus a 5 was produced as a fusion protein with glutathione-S-transferase in Escherichia coli. The recombinant Mus a 5 was purified under native conditions by a combination of affinity, ion-exchange, and reversed phase chromatography. N-terminal sequence was confirmed by Edman degradation and 55 % of the primary structure was identified by mass fingerprint, while the secondary structure was assessed by circular dichroism spectroscopy. IgG reactivity of recombinant protein was shown in 2-D immunoblot with anti-Mus a 5 antibodies, while IgG and IgE binding to natural Mus a 5 was inhibited with the recombinant Mus a 5 in immunoblot inhibition test. IgE reactivity of recombinant Mus a 5 was shown in ELISA within a group of ten persons sensitized to banana fruit. Recombinant Mus a 5 is a novel reagent suitable for the component-resolved allergy diagnosis of banana allergy.

Keywords

Allergen Banana glucanase Food allergy IgE reactivity Mus a 5 

Notes

Acknowledgments

This study was supported by the Ministry of Education, Science and Technological Development, Republic of Serbia, Grant No. 172049. The authors acknowledge support of the FP7 RegPot project FCUB ERA GA No. 256716. The EC does not share responsibility for the content of the article.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Ivan Mrkic
    • 1
  • Mohamed Abughren
    • 1
  • Jasna Nikolic
    • 1
  • Uros Andjelkovic
    • 2
  • Emilia Vassilopoulou
    • 3
  • Athanassios Sinaniotis
    • 3
    • 4
  • Arnd Petersen
    • 5
  • Nikolaos G. Papadopoulos
    • 3
  • Marija Gavrovic-Jankulovic
    • 1
    Email author
  1. 1.Department of Biochemistry, Faculty of ChemistryUniversity of BelgradeBelgradeSerbia
  2. 2.Department of Chemistry, Institute for Chemistry, Technology and MetallurgyUniversity of BelgradeBelgradeSerbia
  3. 3.Allergy Research Centre, 2nd Department of PediatricsUniversity of AthensAthensGreece
  4. 4.Allergy Department“Sotiria” General HospitalAthensGreece
  5. 5.Division Clinical and Molecular AllergologyResearch Center Borstel, Airway Research Center North (ARCN), Member of the German Center for Lung ResearchBorstelGermany

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