Insect Cell-Expressed Major Ragweed Allergen Amb a 1.01 Exhibits Similar Allergenic Properties to Its Natural Counterpart from Common Ragweed Pollen

Maria-Roxana Buzan, Manuela Grijincu, Lauriana-Eunice Zbîrcea, Laura Haidar, Tudor-Paul Tamaș, Monica-Daniela Cotarcă, Gabriela Tănasie, Milena Weber, Elijahu Babaev, Frank Stolz, Rudolf Valenta, Virgil Păunescu, Carmen Panaitescu, Kuan-Wei Chen

Research output: Journal article (peer-reviewed)Journal article

Abstract

Common ragweed pollen allergy has become a health burden worldwide. One of the major allergens in ragweed allergy is Amb a 1, which is responsible for over 90% of the IgE response in ragweed-allergic patients. The major allergen isoform Amb a 1.01 is the most allergenic isoform in ragweed pollen. So far, no recombinant Amb a 1.01 with similar allergenic properties to its natural counterpart (nAmb a 1.01) has been produced. Hence, this study aimed to produce a recombinant Amb a 1.01 with similar properties to the natural isoform for improved ragweed allergy management. Amb a 1.01 was expressed in insect cells using a codon-optimized DNA construct with a removable N-terminal His-Tag (rAmb a 1.01). The recombinant protein was purified by affinity chromatography and physicochemically characterized. The rAmb a 1.01 was compared to nAmb a 1.01 in terms of the IgE binding (enzyme-linked immunosorbent assay (ELISA), immunoblot) and allergenic activity (mediator release assay) in well-characterized ragweed-allergic patients. The rAmb a 1.01 exhibited similar IgE reactivity to nAmb a 1.01 in different IgE-binding assays (i.e., IgE immunoblot, ELISA, quantitative ImmunoCAP inhibition measurements). Furthermore, the rAmb a 1.01 showed comparable dose-dependent allergenic activity to nAmb a 1.01 regarding basophil activation. Overall, the results showed the successful expression of an rAmb a 1.01 with comparable characteristics to the corresponding natural isoform. Our findings provide the basis for an improvement in ragweed allergy research, diagnosis, and immunotherapy.

Original languageEnglish
Article number5175
JournalInternational Journal of Molecular Sciences
Volume25
Issue number10
DOIs
Publication statusPublished - 09 May 2024

Keywords

  • Humans
  • Antigens, Plant/immunology
  • Immunoglobulin E/immunology
  • Animals
  • Allergens/immunology
  • Ambrosia/immunology
  • Recombinant Proteins/immunology
  • Female
  • Adult
  • Plant Proteins/immunology
  • Rhinitis, Allergic, Seasonal/immunology
  • Male
  • Middle Aged
  • Plant Extracts/chemistry

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