Extract-Shaped Immune Repertoires as Source for Nanobody-Based Human IgE in Grass Pollen Allergy.


Journal

Molecular biotechnology
ISSN: 1559-0305
Titre abrégé: Mol Biotechnol
Pays: Switzerland
ID NLM: 9423533

Informations de publication

Date de publication:
Sep 2023
Historique:
received: 06 12 2022
accepted: 11 01 2023
medline: 23 8 2023
pubmed: 26 1 2023
entrez: 25 1 2023
Statut: ppublish

Résumé

The presence of allergen-specific IgE in serum is a biomarker for allergic disease. Specific IgE antibodies for research and diagnostics, however, remain scarce. In contrast to prototypic antibodies, camelid species have evolved single domains as moiety for antigen recognition. These so-called nanobodies represent a versatile platform for the development of diagnostic and therapeutic approaches. In this study, we aimed for generating nanobodies and derived IgE formats from an extract-shaped immune repertoire. Timothy grass pollen represents a complex, but well-defined mixture of individual allergens. Therefore, a repertoire library from a timothy grass pollen extract immunised llama was established. The selection by phage display yielded 3 nanobodies with immunoreactivity to the extract. IgE-like nanobody-based human IgE (nb-hIgE) antibodies were produced in mammalian cells and assessed in different immunoassays and commercial platforms. Immunoblotting and diagnostic ImmunoCap analysis of single timothy grass pollen allergens identified the major allergens Phl p 6 and Phl p 4 as targets. Assessment of immunoreactivity further documented significant molecular cross-reactivity with pollen extract of different grass species and variant presence of allergens within extracts of Pooideae grasses. In summary, our study shows that extract-based immunisation enables the generation of allergen-specific nanobodies and derived nb-hIgE formats linking nanobody technologies with allergological applications.

Identifiants

pubmed: 36696011
doi: 10.1007/s12033-023-00664-8
pii: 10.1007/s12033-023-00664-8
doi:

Substances chimiques

Single-Domain Antibodies 0
Allergens 0
Immunoglobulin E 37341-29-0
Plant Proteins 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

1518-1527

Subventions

Organisme : Novo Nordisk Fonden
ID : 19OC0058484
Organisme : Danmarks Frie Forskningsfond
ID : 9041-00291A

Informations de copyright

© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

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Auteurs

Josephine Baunvig Aagaard (JB)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark.

Michaela Fischer (M)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark.

Julia Lober (J)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark.

Frederikke Bolbro Neumann (FB)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark.

Dena Allahverdi (D)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark.

Coline Sivelle (C)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark.

Michaela Miehe (M)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark.

Edzard Spillner (E)

Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Gustav Wieds Vej 10, 8000, Aarhus C, Denmark. e.spillner@bce.au.dk.

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