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

Background: Pemphigus vulgaris (PV) is a life-threatening autoimmune blistering disease caused mainly by IgG autoantibodies (auto-abs) against the cadherintype adhesion molecules desmoglein (Dsg) 1 and 3. Pathogenic anti-Dsg3 autoabs bind to different Dsg3 epitopes, leading, among others, to sign...

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Glavni autori: Eming, Rüdiger, Riaz, Shafaq, Müller, Eliane J., Zakrzewicz, Anna, Linne, Uwe, Tikkanen, Ritva, Zimmer, Christine Lea, Hudemann, Christoph
Format: Članak
Jezik:engleski
Izdano: Philipps-Universität Marburg 2024
Teme:
autoimmunity
2G4
antibody
quality control
pemphigus vulgaris
Medizin
PV
desmoglein (Dsg)
Medical sciences Medicine
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Opis
Sažetak:Background: Pemphigus vulgaris (PV) is a life-threatening autoimmune blistering disease caused mainly by IgG autoantibodies (auto-abs) against the cadherintype adhesion molecules desmoglein (Dsg) 1 and 3. Pathogenic anti-Dsg3 autoabs 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 antigenspecific 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.
Opis predmeta:Gefördert durch den Open-Access-Publikationsfonds der UB Marburg.
Digitalni identifikator objekta:10.3389/fimmu.2024.1464881

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