Quality-controlled characterization of a monoclonal antibody specific to an EC5-domain of human desmoglein 3 for pemphigus research.

Autor:	Eming R; Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany.; Department of Dermatology, Venerology and Allergology, German Armed Forces Central Hospital Koblenz, Koblenz, Germany., Riaz S; Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany., Müller EJ; Department for BioMedical Research, Molecular Dermatology and Stem Cell Research, University of Bern, Bern, Switzerland.; Department of Dermatology, Inselspital, Bern University Hospital, University of Bern, Bern, Switzerland., Zakrzewicz A; Institute of Biochemistry, Medical Faculty, Justus-Liebig-University Giessen, Giessen, Germany., Linne U; Mass Spectrometry Facility, Department of Chemistry, Philipps University, Marburg, Germany., Tikkanen R; Institute of Biochemistry, Medical Faculty, Justus-Liebig-University Giessen, Giessen, Germany., Zimmer CL; Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany., Hudemann C; Department of Dermatology and Allergology, Philipps University Marburg, Marburg, Germany.
Jazyk:	angličtina
Zdroj:	Frontiers in immunology [Front Immunol] 2024 Oct 10; Vol. 15, pp. 1464881. Date of Electronic Publication: 2024 Oct 10 (Print Publication: 2024).
DOI:	10.3389/fimmu.2024.1464881
Abstrakt:	Background: 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. Methods: 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. Results: 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. Conclusion: 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. Competing Interests: RE is recipient of an unrestricted grant from Topas Therapeutics, Hamburg, Germany. RT has received funding from argenx. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be constructed as a potential conflict of interest. (Copyright © 2024 Eming, Riaz, Müller, Zakrzewicz, Linne, Tikkanen, Zimmer and Hudemann.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu