Characterization of monoclonal ZnT8-specific antibody

Abstract

Event Abstract Back to Event Characterization of monoclonal ZnT8-specific antibody Julia Ustinova1*, Denis Belitskin2, Erkki Juronen3, Eva Zusinaite4, Meeme Utt1 and Raivo Uibo1 1 Institute of Biomedicine, University of Tartu, Estonia 2 Institute of Molecular and Cell Biology, University of Tartu, Estonia 3 Department of Biomedicine, University of Tartu, Estonia 4 Institute of Technology, University of Tartu, Estonia Immune-mediated destruction of insulin-producing beta cells of pancreas leads to autoimmune type 1 diabetes mellitus (T1D). Recently, ZnT8 (zinc transporter 8) has been identified as an autoimmune target in T1D. ZnT8 is expressed in the pancreatic tissue and regulates Zn metabolism in beta cells. Zn is critical for normal storage of insulin in the secretory vesicles of islet beta cells. ZnT8 autoantibodies are directed mostly against the C terminal domain. Molecular biological tools for basic research of ZnT8 role in T1D are needed. In this report, we characterize a new monoclonal antibody to the ZnT8 C-terminus. Mice were immunized with Semliki Forest virus-like particles expressing full-length ZnT8 and hybridomas were obtained and cloned. Hybridomas were tested using LIPS (Luciferase Immunoprecipitation system) and ELISA using purified recombinant C-terminal domain of ZnT8. One of the hybridomas giving a very high signal was selected for further analysis. Protein A purified Mab was used to characterize binding epitope using 18 overlapping peptides designed according to C-terminal sequence of ZnT8. Binding epitope was identified using four methods: LIPS assay, ELISA, biosensor binding test, and immunohistochemistry assay with monkey pancreatic iselets. In conclusion, we have characterized a new monoclonal antibody usable as a molecular biological tool for ZnT8 research. Keywords: ZnT8, Autoantibodies, autoimmune diabetes, Monoclonal antibody, beta cells Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Immune-mediated disease pathogenesis Citation: Ustinova J, Belitskin D, Juronen E, Zusinaite E, Utt M and Uibo R (2013). Characterization of monoclonal ZnT8-specific antibody. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00326 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 14 Mar 2013; Published Online: 22 Aug 2013. * Correspondence: Miss. Julia Ustinova, Institute of Biomedicine, University of Tartu, Tartu, Estonia, lia@ut.ee Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Julia Ustinova Denis Belitskin Erkki Juronen Eva Zusinaite Meeme Utt Raivo Uibo Google Julia Ustinova Denis Belitskin Erkki Juronen Eva Zusinaite Meeme Utt Raivo Uibo Google Scholar Julia Ustinova Denis Belitskin Erkki Juronen Eva Zusinaite Meeme Utt Raivo Uibo PubMed Julia Ustinova Denis Belitskin Erkki Juronen Eva Zusinaite Meeme Utt Raivo Uibo Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.