Isolation of nanobodies with potential to reduce patients' IgE binding to Bet v 1
| Autor: | Christina Weichwald, Barbara Bohle, Maria R. Strobl, Sergei Tillib, Ines Zettl, Evgenia Khan, Oksana Goryainova, Margarete Focke-Tejkl, Sabine Flicker, Anja Drescher, Tatiana B. Ivanova, Marina V. Rutovskaya |
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| Rok vydání: | 2021 |
| Předmět: |
Phage display
medicine.drug_class Immunology chemical and pharmacologic phenomena Immunoglobulin E Monoclonal antibody Epitope law.invention Epitopes law medicine Hypersensitivity Immunology and Allergy Humans Plant Proteins biology Chemistry hemic and immune systems Allergens Antigens Plant Single-Domain Antibodies Molecular biology Basophil activation Polyclonal antibodies biology.protein Recombinant DNA Pollen Antibody |
| Zdroj: | AllergyREFERENCES. 77(6) |
| ISSN: | 1398-9995 |
| Popis: | BACKGROUND Recent studies showed that a single injection of human monoclonal allergen-specific IgG antibodies significantly reduced allergic symptoms in birch pollen-allergic patients. Since the production of full monoclonal antibodies in sufficient amounts is laborious and expensive, we sought to investigate if smaller recombinant allergen-specific antibody fragments, i.e. nanobodies, have similar protective potential. For this purpose, nanobodies specific for Bet v 1, the major birch pollen allergen, were generated to evaluate their efficacy to inhibit IgE-mediated responses. METHODS A cDNA-VHH library was constructed from a camel immunized with Bet v 1 and screened for Bet v 1 binders encoding sequences by phage display. Selected nanobodies were expressed, purified and analyzed in regards of epitope-specificity and affinity to Bet v 1. Furthermore, cross-reactivity to Bet v 1-homologues from alder, hazel and apple, and their usefulness to inhibit IgE binding and allergen-induced basophil activation were investigated. RESULTS We isolated three nanobodies that recognize Bet v 1 with high affinity and cross-react with Aln g 1 (alder) and Cor a 1 (hazel). Their epitopes were mapped to the alpha-helix at the C-terminus of Bet v 1. All nanobodies inhibited allergic patients´ polyclonal IgE binding to Bet v 1, Aln g 1 and Cor a 1 and partially suppressed Bet v 1-induced basophil activation. CONCLUSION We identified high-affinity Bet v 1-specific nanobodies that recognize an important IgE-epitope and reduce allergen-induced basophil activation revealing the first proof that allergen-specific nanobodies are useful tools for future treatment of pollen allergy. |
| Databáze: | OpenAIRE |
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