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

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. © 2011 bentham science publishers ltd.