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

Author:

Buzan Maria-Roxana12ORCID,Grijincu Manuela12ORCID,Zbîrcea Lauriana-Eunice12ORCID,Haidar Laura1ORCID,Tamaș Tudor-Paul1ORCID,Cotarcă Monica-Daniela1,Tănasie Gabriela12,Weber Milena3,Babaev Elijahu4,Stolz Frank4,Valenta Rudolf3567,Păunescu Virgil12,Panaitescu Carmen12ORCID,Chen Kuan-Wei2

Affiliation:

1. Center of Immuno-Physiology and Biotechnologies, Department of Functional Sciences, Victor Babes University of Medicine and Pharmacy, 300041 Timisoara, Romania

2. OncoGen Center, Pius Brinzeu County Clinical Emergency Hospital, 300723 Timisoara, Romania

3. Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, 1090 Vienna, Austria

4. Vienna Competence Center, Biomay AG, 1090 Vienna, Austria

5. Laboratory for Immunopathology, Department of Clinical Immunology and Allergology, Sechenov First Moscow State Medical University, 119991 Moscow, Russia

6. Karl Landsteiner University of Health Sciences, 3500 Krems, Austria

7. NRC Institute of Immunology FMBA of Russia, 115478 Moscow, Russia

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.

Funder

INSPIRED project

Romanian Society of Allergology and Clinical Immunology

Danube Allergy Research Cluster of the Country of Lower Austria

Russian Science Foundation

Victor Babes University of Medicine and Pharmacy Timisoara

Publisher

MDPI AG

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