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-Jankulovic

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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.

Original languageEnglish
Pages (from-to)498-506
Number of pages9
JournalMolecular Biotechnology
Volume56
Issue number6
DOIs
Publication statusPublished - 2014

Keywords

  • Allergen
  • Banana glucanase
  • Food allergy
  • IgE reactivity
  • Mus a 5

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    Mrkic, I., Abughren, M., Nikolic, J., Andjelkovic, U., Vassilopoulou, E., Sinaniotis, A., Petersen, A., Papadopoulos, N. G., & Gavrovic-Jankulovic, M. (2014). Molecular characterization of recombinant Mus a 5 allergen from banana fruit. Molecular Biotechnology, 56(6), 498-506. https://doi.org/10.1007/s12033-013-9719-8