An Ultrasensitive, High-Throughput, and Low-Cost Autoimmunity Panel for the Early Diagnosis and Improved Treatment of Type 1 Diabetes

1型糖尿病（T1D）是一种破坏性的自身免疫性疾病，破坏胰岛细胞胰岛素的产生。数以百万计的美国人，特别是儿童，受到影响，必须终身依赖胰岛素治疗，并与危及生命的合并症作斗争。大多数T1D新病例是在紧急情况下诊断的，表现为潜在致命的糖尿病酮症酸中毒（DKA）。目前可以对T1D采取预防行动的临床前诊断测试速度缓慢，而且在三级保健中心之外不易获得。在这项提案中，Enable Biosciences将进一步开发和验证一种新的低成本、高通量的免疫测定方法，以快速、灵敏地筛选预测1型糖尿病风险的四种主要自身抗体标记物。这样的测试可以取代目前昂贵、缓慢的放射性方法。通过降低成本，同时保持极低样本量检测的灵敏度和特异性，Enable Biosciences的产品将允许增加检测，在最佳干预时间发现普通人群中的早期T1D。在I期融资期间，Enable利用其核心抗体检测技术ADAP （Antibody Detection by Agglutination-PCR）制作了一个3成员T1D自身抗体面板的原型，并证明了与金标准分析的基本等效性，同时消除了对放射性试剂的依赖。在第二阶段提案中，Enable提出了一个全面的计划，通过使用专用机器人液体处理系统的自动化来增强该面板，大大提高了分析通量和可靠性，同时进一步降低了成本。我们还将调整我们的分析化学，以便能够使用Taqman探针。这将导致进一步的吞吐量加速和成本降低。基于第一阶段工作的初步结果，我们将纳入第四种也是最后一种T1D自身抗体（ZnT8），以创建世界上第一个“4合1”筛查面板，创建一个为T1D筛查和诊断提供最高水平风险评估的产品。这些优化和自动化的面板将通过我们广泛的热情的学术和机构合作伙伴网络提供的数千个临床样本进行严格验证，这些合作伙伴包括乔斯林糖尿病中心，斯坦福医学院，T1D交流和塔夫茨儿童医院。这些数据将用于规划和指导商业化工作，包括分阶段的方法，从提供clia认证的实验室服务到fda批准的试剂盒，以及配套的样品到答案的集成硬件设备，以便在全球的研究和临床实验室和筛选地点轻松，低技能地使用和部署。

Type 1 diabetes (T1D) is a devastating autoimmune disease that destroys islet-cell insulin production. Millions of Americans, especially children, are affected and must rely on lifelong insulin administration and struggle with life-threatening co-morbidities. Most new cases of T1D are diagnosed in emergency settings with presentation in potentially deadly diabetic ketoacidosis (DKA). Current pre-clinical diagnostic tests that can permit preventive action for T1D are slow and not readily available outside of tertiary care centers. In this proposal Enable Biosciences will further develop and validate a novel low-cost and high-throughput immunoassay to rapidly and sensitively screen for the four cardinal autoantibody markers that predict risk for type 1 diabetes. Such a test could supplant the current expensive, slow radioactive method. By reducing costs while retaining sensitivity and specificity with a robust very low-sample volume assay, Enable Biosciences' product will permit increased testing to reveal incipient T1D in the general population at the optimal time for intervention. During the Phase I funding period, Enable prototyped a 3-member T1D autoantibody panel using their core Antibody Detection by Agglutination-PCR (ADAP) technology and demonstrated substantial equivalence with the gold-standard assay while removing reliance on radioactive reagents. In this Phase 2 proposal, Enable presents a comprehensive plan to enhance this panel via automation using a dedicated robotic liquid handling system, greatly increasing assay throughput and reliability while further reducing costs. We will also adapt our assay chemistry to be able to use Taqman probes. This will result in further throughput acceleration and cost reductions. Building upon promising preliminary results from Phase 1 work, we will incorporate the fourth and final T1D autoantibody (ZnT8) to create the world's first ever “4-in-1” screening panel, creating a product that provides the highest level of risk assessment for T1D screening and diagnosis. These optimized and automated panels will then be rigorously validated with thousands of clinical samples provided by our wide network of enthusiastic academic and institutional partners that include the Joslin Diabetes Center, Stanford School of Medicine, the T1D Exchange and Tufts Children's Hospital. This data will be used to plan and direct commercialization efforts that will include a phased approach from delivering a CLIA-certified laboratory service to FDA-approved reagent kits with companion integrated sample-to-answer hardware devices for easy, low-skill use and deployment across research and clinical laboratories and screening sites worldwide.