The following proposal for the purchase of a Sector Imager 6000 Electrochemiluminescence plate reader is directed towards developing a high throughput shared facility for the analysis of immunological biomarkers of autoimmune disease and in particular autoantibodies and cytokines that play a role in the diagnosis and pathogenesis of type 1 autoimmune diabetes in humans. The Barbara Davis Center for Childhood Diabetes, where the instrument will be housed, is one of six reference laboratories in the world for the analysis of autoantibodies in new-diagnosed patients and for the identification of autoantibody positive individuals at risk of developing disease. The latter have been recruited into numerous national and international translational research studies following the natural history of the disease (DPT-1, TrialNet) and trails of experimental therapies (recently MMF, Rituximab, Bayhill). The SECTOR(R) Imager 6000 is extremely adaptable to a multiuser, multi-assay format without the need for calibration, resetting instrument parameters as well as being easily accessible compared to instruments such as scintillation counters and flow cytometers that are used in many of the current assays. It is now possible through the use of autoantibody testing combined with genetic analysis to identify individuals at risk of developing the disease from the general population however to achieve this goal there is a need to implement autoantibody assays using new formats that do not use radioisotopes yet can provide quantitative data that can be reproduced in multiple laboratories in very different operating environments all over the world. The MSD technology we contend is an excellent choice in that it brings us to the forefront of technology, yet provides a platform that is versatile and can be used for research and development of new assays and at the same time will be transplantable to fully-automated lab-on-a-chip assays that might ultimately be used as a point of service assay. Electrochemiluminescence detection affords distinct advantages in terms of speed, signal to noise and wide dynamic range over radiometric and photometric technologies. The ability to multiplex up to 10 analytes per well on prespotted plates is a significant saving in terms of reagents and operator time. The following proposal identifies a group of 4 major users involved in translational research and routine assay performance (70% instrument use) and 7 minor users (30% instrument use) who will principally use the machine for experimental research in animal models and in the research and development of new biomarkers of disease. Support from 20 NIH funded grants is documented ranging from several multicenter consortia to DERC pilot and feasibility grant holders. A plan for management and maintenance of the instrument is provided. Validation and quality control of the assays will be performed through our existing participation in the Diabetes Autoimmunity Standardization Program run from the Center of Disease control through the Immunology of Diabetes Society.
