Evaluation of different glycoforms of honeybee venom major allergen phospholipase A2 (Api m 1) produced in insect cells

Simon Blank, Yvonne Michel, Henning Seismann, Melanie Plum, Kerstin Greunke, Thomas Grunwald, Reinhard Bredehorst, Markus Ollert, Ingke Braren, Edzard Spillner*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

33 Citations (Scopus)

Abstract

Allergic reactions to hymenoptera stings are one of the major reasons for IgE-mediated anaphylaxis. However, proper diagnosis using venom extracts is severely affected by molecular cross-reactivity. In this study recombinant honeybee venom major allergen phospholipase A2 (Api m 1) was produced for the first time in insect cells. Using baculovirus infection of different insect cell lines allergen versions providing a varying degree of cross-reactive carbohydrate determinants as well as a non glycosylated variant could be obtained as secreted soluble proteins in high yields. The resulting molecules were analyzed for their glycosylation and proved to show advantageous properties regarding cross-reactivity in sIgE-based assays. Additionally, in contrast to the enzymatically active native protein the inactivated allergen did not induce IgE-independent effector cell activation. Thus, insect cell-derived recombinant Api m 1 with defined CCD phenotypes might provide further insights into hymenoptera venom IgE reactivities and contribute to an improved diagnosis of hymenoptera venom allergy.

Original languageEnglish
Pages (from-to)415-422
Number of pages8
JournalProtein and Peptide Letters
Volume18
Issue number4
DOIs
Publication statusPublished - Apr 2011
Externally publishedYes

Keywords

  • Allergen
  • Baculovirus
  • Carbohydrates
  • Cross-reactivity
  • Honeybee
  • Phospholipase
  • Recombinant allergen
  • Venom allergy

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