Purification and characterization of natural Bet v 1 from birch pollen and related allergens from carrot and celery

M.A. Bollen, A. Garcia, J.H.G. Cordewener, H.J. Wichers, J.P.F.G. Helsper, H.F.J. Savelkoul, M.A.J.S. van Boekel

Research output: Contribution to journalArticleAcademicpeer-review

30 Citations (Scopus)

Abstract

Birch pollen allergy is predominantly caused by the major allergen Bet v 1 and can lead to crossreactions with homologous proteins in food. Two major cross-reactive food allergens are Dau c 1 from carrot and Api g 1 from celery, which have never been purified from their natural source. Here, we describe a non-denaturing purification method for obtaining natural Bet v 1, Dau c 1 and Api g 1, comprising of ammonium sulfate precipitation, hydrophobic interaction chromatography and size exclusion chromatography. This method resulted in 98-99% pure isoform mixtures for each allergen. Characterization of these isoform mixtures with Q-TOF MS/MS clearly showed earlier reported isoforms of Bet v 1, Dau c 1 and Api g 1, but also new isoforms. The presence of secondary structure in the three purified allergens was demonstrated via circular dichroism and showed high similarity. The immune reactivity of the natural allergens was compared with recombinant proteins by Western blot and ELISA and showed similar reactivity.
Original languageEnglish
Pages (from-to)1527-1536
JournalMolecular Nutrition & Food Research
Volume51
Issue number12
DOIs
Publication statusPublished - 2007

Keywords

  • low-temperature method
  • major cherry allergen
  • bet-v-i
  • cross-reactivity
  • molecular characterization
  • mass-spectrometry
  • apium-graveolens
  • apple allergen
  • plant-tissues
  • pru av-1

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