WEBINAR: High Throughput Production of Autoantigens for Rapid Detection of Multiple Autoimmune Diseases

Speaker
  • Professor in Medicine, Professor in Pharmacology and Pharmacy, and Director of State Key Laboratory of Pharmaceutical Biotechnology, University of Hong Kong
    Biography
      Dr. Aimin Xu is currently a chair professor for the Department of Medicine and the Department of Pharmacology & Pharmacy and the director of the State Key Laboratory of Pharmaceutical Biotechnology at the University of Hong Kong. His major research interest is the discovery and functional characterization of novel hormones and biomarkers involved in the pathogenesis of metabolic and autoimmune diseases. His team has identified the glycol-isoforms of adiponectin and the FGF21-adiponectin axis in protection against obesity-related cardiometabolic syndrome. His team also discovered adipocyte fatty acid binding protein (A-FABP) and lipocalin-2 as pro-inflammatory adipokines in both rodents and humans. In addition, his work contributed significantly to the understanding of the molecular basis of adipose tissue inflammation and immunometabolic regulators in both metabolic and autoimmune diseases. His team has developed a series of immunoassays that are now widely used for clinical diagnostics, drug screening, and clinical and basic research for metabolic and immunological disorders

    Abstract

    Current methods for in vitro diagnosis of autoimmune diseases (ADs) are mainly based on the detection of circulating autoantibodies using specific AD autoantigens. A high-quality recombinant autoantigen with epitopes resembling its natural form in the human body is the key to sensitive, specific, and rapid diagnosis. In this webinar we describe the use of the Gibco™ Bac-to-Bac™ Baculovirus Expression System to express 22 recombinant autoantigens in Spodoptera frugiperda 9 (Sf9) cells, with folding and posttranslational modifications matching those of the natural autoantigens in vivo. The Bac-to-Bac system allowed us to generate recombinant baculoviruses expressing the target genes via transposition within 48 hours. Using Gibco™ Sf-900™ II SFM, we were able to propagate Sf9 cells to a density of approximately 6–8 x 106 cells/mL in a suspension culture and over 1–2 x 107 cells/mL during fermentation. These tools significantly reduce the time and labor costs needed for virus generation, amplification, and protein production, and allow high-throughput production of autoantigens. Using these 22 high-quality recombinant autoantigens, we have successfully developed a membrane-based chip for simultaneous screening of 7 major ADs that were validated in over 200 clinical samples. In addition to demonstrating greater than 95% sensitivity, the new assay procedure is very rapid (only taking 30 minutes).
        
    Learning Objectives:

    • Understand the advantages of the Bac-to-Bac Baculovirus Expression System
    • Demonstrate how the Bac-to-Bac Baculovirus Expression System can be used for high throughput production of high-quality recombinant autoantigens

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