Open Peer Review Discuss this article (0) Comments ANTIBODY VALIDATION ARTICLE Optimized purification strategies for the elimination of non-specific products in the isolation of GAD65-specific monoclonal autoantibodies [version 2; referees: 2 approved, 1 approved with reservations] Wei Jiang , Henriette Macmillan , Anne-Marie Madec , Elizabeth D. Mellins 1,2 Department of Pediatrics, Stanford University, Stanford, CA, 94305, USA Stanford Program in Immunology, Stanford University, Stanford, CA, 94305, USA Department of Medicine, University of California San Francisco, San Francisco, CA, 94143, USA INSERM U1060, Faculté de médecine Lyon-Sud, Oullins Cedex, France Abstract Autoantibodies against antigens expressed by insulin-producing β cells are circulating in both healthy individuals and patients at risk of developing Type 1 diabetes. Recent studies suggest that another set of antibodies (anti-idiotypic antibodies) exists in this antibody/antigen interacting network to regulate auto-reactive responses. Anti-idiotypic antibodies may block the antigen-binding site of autoantibodies or inhibit autoantibody expression and secretion. The equilibrium between autoantibodies and anti-idiotypic antibodies plays a critical role in mediating or preventing autoimmunity. In order to investigate the molecular mechanisms underlying such a network in autoimmunity and potentially develop neutralizing reagents to prevent or treat Type 1 diabetes, we need to produce autoantibodies and autoantigens with high quality and purity. Herein, using GAD65/anti-GAD65 autoantibodies as a model system, we aimed to establish reliable approaches for the preparation of highly pure autoantibodies suitable for downstream investigation. 1,2 1-3 4 1,2 1 2 3 4 Referee Status: Invited Referees version 2 published 21 Apr 2016 version 1 published 29 May 2015 1 2 3 report report report , University of Iowa, USA David Soll 1 , University of Utah, USA Xiao He 2 , University of Christiane Hampe Washington, USA 3 29 May 2015, :135 (doi: ) First published: 4 10.12688/f1000research.6467.1 21 Apr 2016, :135 (doi: ) Latest published: 4 10.12688/f1000research.6467.2 v2 This article is included in the Antibody Validations SPONSORED GATEWAY Organizations supporting this gateway have had no editorial control with respect to the content contained herein. Page 1 of 13 F1000Research 2016, 4:135 Last updated: 03 JUL 2017