A Novel Method for Total IgE Purification from Human Serum.


Journal

Journal of immunology (Baltimore, Md. : 1950)
ISSN: 1550-6606
Titre abrégé: J Immunol
Pays: United States
ID NLM: 2985117R

Informations de publication

Date de publication:
15 05 2022
Historique:
received: 26 05 2021
accepted: 17 03 2022
pubmed: 7 5 2022
medline: 20 5 2022
entrez: 6 5 2022
Statut: ppublish

Résumé

For Ab purification, high-affinity chromatography is commonly used. This technique results in high-purity Abs, but it requires highly specific knowledge and equipment. Commercial kits for purification of IgE are not available. Therefore, we established a (to our knowledge) novel method for the purification of total IgE from human serum. Sera from 19 allergic and nonallergic patients were included. After depletion of polyclonal IgG, total serum IgE was captured using anti-human IgE Abs coupled to beads, eluted from the beads, and incubated with protein G-coupled beads to increase the final purity. Purity analysis and Ab detection were performed by Western blot. Total serum IgE and purified IgE concentrations were analyzed using ELISA. To determine their functionality, primary human mast cells were sensitized with purified IgE and activated with anti-IgE or a relevant allergen. CD63

Identifiants

pubmed: 35523453
pii: jimmunol.2100497
doi: 10.4049/jimmunol.2100497
doi:

Substances chimiques

Allergens 0
Immunoglobulin G 0
Immunoglobulin E 37341-29-0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

2436-2442

Informations de copyright

Copyright © 2022 by The American Association of Immunologists, Inc.

Auteurs

Fariza Mishaal Saiema Badloe (FMS)

Vrije Universiteit Brussel (VUB), SKIN Research Group, Brussels, Laarbeeklaan 103, 1090 Belgium.
Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Department of Dermatology, Laarbeeklaan 101, 1090 Brussels, Belgium.

Shauni De Vriese (S)

Vrije Universiteit Brussel (VUB), SKIN Research Group, Brussels, Laarbeeklaan 103, 1090 Belgium.
Institute of Allergology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.

Tina De Bruyn Carlier (T)

Vrije Universiteit Brussel (VUB), SKIN Research Group, Brussels, Laarbeeklaan 103, 1090 Belgium.

Elise Vandersichel (E)

Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Department of Dermatology, Laarbeeklaan 101, 1090 Brussels, Belgium.

Jörg Scheffel (J)

Institute of Allergology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
Fraunhofer Institute for Translational Medicine and Pharmacology, Allergology and Immunology, Berlin, Germany; and.

Marcus Maurer (M)

Institute of Allergology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
Fraunhofer Institute for Translational Medicine and Pharmacology, Allergology and Immunology, Berlin, Germany; and.

Johannes Ring (J)

Department of Dermatology and Allergology Biederstein, Technical University Munich, Munich, Germany.

Jan Gutermuth (J)

Vrije Universiteit Brussel (VUB), SKIN Research Group, Brussels, Laarbeeklaan 103, 1090 Belgium.
Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Department of Dermatology, Laarbeeklaan 101, 1090 Brussels, Belgium.

Inge Kortekaas Krohn (I)

Vrije Universiteit Brussel (VUB), SKIN Research Group, Brussels, Laarbeeklaan 103, 1090 Belgium; ingekortekaas@gmail.com.
Vrije Universiteit Brussel (VUB), Universitair Ziekenhuis Brussel (UZ Brussel), Department of Dermatology, Laarbeeklaan 101, 1090 Brussels, Belgium.

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Classifications MeSH