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ORIGINAL RESEARCH article
Front. Immunol.
Sec. Immunological Tolerance and Regulation
Volume 15 - 2024 |
doi: 10.3389/fimmu.2024.1464881
This article is part of the Research Topic Community Series in Autoantibodies: Volume II View all 7 articles
Quality-controlled characterization of a monoclonal antibody specific to an EC5-domain of human desmoglein 3 for pemphigus research
Provisionally accepted
Rüdiger Eming
1,2*
Shafaq Riaz
1*
Eliane Müller
3
Anna Zakrzewicz
4
Uwe Linne
5
Christine L. Zimmer
1*
Ritva Tikkanen
4
Christoph Hudemann
1*- 1 Department of Dermatology and Allergology, Philipps-University Marburg, Marburg, Germany
- 2 Department of Dermatology, Venerology and Allergology, German Armed Forces Central Hospital Koblenz, Koblenz, Germany
- 3 Department for BioMedical Research, Molecular Dermatology and Stem Cell Research, University of Bern, Bern, Switzerland
- 4 Institute of Biochemistry, Medical Faculty, Justus-Liebig-University Giessen, Giessen, Germany
- 5 Mass Spectrometry Facility, Department of Chemistry, Philipps University Marburg, Marburg, Germany
Pemphigus vulgaris (PV) is a life-threatening autoimmune blistering disease caused mainly by IgG autoantibodies (auto-abs) against the cadherin-type adhesion molecules desmoglein (Dsg) 1 and 3. Pathogenic anti-Dsg3 auto-abs bind to different Dsg3 epitopes, leading, among others, to signalling that is involved in pathogenic events, such as Dsg3 depletion. As central tools in research on PV, a limited number of antibodies such as AK23 are frequently used by the autoimmune bullous disease community. Previously, we have introduced a novel Dsg3 EC5-binding antibody termed 2G4 that may potentially serve as a superior tool for numerous PV related analysis. The purpose of this study was to develop a quality-controlled production and verification process that allows I) a continuous quality improvement, and II) a verified and comprehensible overall quality with regard to pathogenic antigen-specific binding in a variety of pemphigus assays for each batch production. Thus, a workflow based on a standardized operating procedure was established. This includes the verification of purity and in-vitro binding capacity (SDS-page, direct and indirect immunofluorescence) as primary parameters, and size analysis by mass-spectrometry and ex-vivo pathogenicity by monolayer dissociation assay. We here present an extensive point-by-point quality controlled IgG production protocol, which will serve as a basis for a standardized antibody assessment in PV research.
Keywords: Quality control, antibody, Pemphigus Vulgaris, Autoimmunity, Desmoglein (Dsg)
Received: 15 Jul 2024; Accepted: 13 Sep 2024.
Copyright: © 2024 Eming, Riaz, Müller, Zakrzewicz, Linne, Zimmer, Tikkanen and Hudemann. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Rüdiger Eming, Department of Dermatology and Allergology, Philipps-University Marburg, Marburg, Germany
Shafaq Riaz, Department of Dermatology and Allergology, Philipps-University Marburg, Marburg, Germany
Christine L. Zimmer, Department of Dermatology and Allergology, Philipps-University Marburg, Marburg, Germany
Christoph Hudemann, Department of Dermatology and Allergology, Philipps-University Marburg, Marburg, Germany
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