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Jiang W, Macmillan H, Madec AM, et al. Optimized purification strategies for the elimination of non-specific products in the isolation of GAD65-specific monoclonal autoantibodies. F1000Res. 2015;4:135doi: 10.12688/f1000research.6467.2.
Jiang, W., Macmillan, H., Madec, A. M., & Mellins, E. D. (2015). Optimized purification strategies for the elimination of non-specific products in the isolation of GAD65-specific monoclonal autoantibodies. F1000Research, 4135. https://doi.org/10.12688/f1000research.6467.2
Jiang, Wei, et al. "Optimized purification strategies for the elimination of non-specific products in the isolation of GAD65-specific monoclonal autoantibodies." F1000Research vol. 4 (2015): 135. doi: https://doi.org/10.12688/f1000research.6467.2
Jiang W, Macmillan H, Madec AM, Mellins ED. Optimized purification strategies for the elimination of non-specific products in the isolation of GAD65-specific monoclonal autoantibodies. F1000Res. 2015 May 29;4:135. doi: 10.12688/f1000research.6467.2. eCollection 2015. PMID: 29167731; PMCID: PMC5680538.
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F1000Res. 2015 May 29;4:135. doi: 10.12688/f1000research.6467.2. eCollection 2015.

Optimized purification strategies for the elimination of non-specific products in the isolation of GAD65-specific monoclonal autoantibodies.

F1000Research

Wei Jiang, Henriette Macmillan, Anne-Marie Madec, Elizabeth D Mellins

Affiliations

  1. Department of Pediatrics, Stanford University, Stanford, CA, 94305, USA.
  2. Stanford Program in Immunology, Stanford University, Stanford, CA, 94305, USA.
  3. Department of Medicine, University of California San Francisco, San Francisco, CA, 94143, USA.
  4. INSERM U1060, Faculté de médecine Lyon-Sud, Oullins Cedex, France.

PMID: 29167731 PMCID: PMC5680538 DOI: 10.12688/f1000research.6467.2

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.

Keywords: GAD65-specific; affinity purification; autoantibody production; monoclonal autoantibody

Conflict of interest statement

Competing interests: No competing interests were disclosed.

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