Evaluation of Different Glycoforms of Honeybee Venom Major Allergen Phospholipase A2 (Api m 1) Produced in Insect Cells
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.
Keywords: Allergen; Allergen Phospholipase A2; CCD phenotypes; Glycoforms; HBV acid phosphatase; Honeybee Venom; IgE-mediated anaphylaxis; MUXF-HAS; RBL-SX38; Recombinant Baculovirus; Site Directed Mutagenesis; agglutinin; baculovirus; carbohydrates; cross-reactivity; honeybee; hymenoptera stings; hymenoptera venom; phospholipase; recombinant allergen; venom allergy
Document Type: Research Article
Publication date: 2011-04-01
- Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.