SBIR Phase I: Development of an ultrasensitive, high-throughput autoantibody discovery platform using agglutination-PCR

小型企业创新研究(SBIR)项目更广泛的影响/商业潜力是开发一种工具，加速发现新的自身抗体生物标记物，用于人类疾病的早期发现和个性化治疗。自身抗体生物标记物被广泛用于诊断各种疾病，如自身免疫性疾病、传染病和癌症。然而，目前自身抗体分析方法的有限的重复性、成本和敏感性阻碍了发现新的生物标记物来改善对这些疾病的管理。一个具有成本效益、高度敏感和强大的自身抗体分析平台可以揭示目前方法可能遗漏的自身抗体谱系中的变化。这种工具将成为发现新的生物标志物的宝贵渠道，增强检测、治疗和了解多种疾病的能力。在每年74亿美元的生物标记物发现阵列市场上，建议的工具以服务或微阵列试剂盒的形式出售供客户使用，满足了几个客户的必备需求，包括寻求新的疾病靶点的生物制药公司、改进临床试验和寻求更好地了解人类生物学的学术团体。这个SBIR第一阶段项目建议开发一种高度多元化和灵敏的自身抗体生物标记物分析工具，以加快生物标记物的发现。提出的方法使用了凝集-聚合酶链式反应，这是加州大学伯克利分校和斯坦福大学开发的一项新技术。凝集-聚合酶链式反应在溶液阶段检测自身抗体，以确保抗原探针的正确折叠，同时利用标准定量聚合酶链式反应仪器的灵敏度和多重能力。该项目采用一种创新的合成策略，以低成本准备和优化一大批探头。虽然传统的合成路线需要长达3个月的时间，但拟议的策略可以将时间减少到一周。此外，用于检测自身抗体的探针库将使用来自健康患者或系统性红斑狼疮(SLE)患者的银行血清/血浆进行验证。这项实验将作为一个强大的原理证明，因为SLE显示了许多不同的自身抗体，用其他方法检测是有挑战性的。建议产品的分析灵敏度和重现性将与标准蛋白质微阵列进行比较。将对该平台进行测试，以确保可重复性和易用性。探针库和方案将作为最低限度可行的产品和在其他疾病中应用的原型发挥作用。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is the development of a tool to accelerate the discovery of new autoantibody biomarkers for the early detection and personalized treatment of human diseases. Autoantibody biomarkers are broadly used to diagnose a variety of conditions, such as autoimmune disorders, infectious diseases and cancers. However, the limited reproducibility, cost and sensitivity of current autoantibody profiling methods frustrate the discovery of new biomarkers to improve management of these diseases. A cost-effective autoantibody profiling platform that is highly sensitive and robust could reveal changes in the autoantibody repertoire that might have been missed by current methods. Such a tool would serve as an invaluable pipeline for the discovery of new biomarkers, augmenting the ability to detect, treat and understand numerous diseases. In the $7.4 Billion/yr biomarker discovery array market, the proposed tool, sold as a service or as micro-arrays kits for client use, meets must-have needs of several customers, including biopharmaceutical companies seeking for new disease targets, improving clinical trials and academic groups pursuing better understanding of human biology.This SBIR Phase I project proposes to develop a highly multiplexed and sensitive autoantibody biomarker profiling tool to expedite biomarker discovery. The proposed approach uses agglutination-PCR, a novel technique developed at UC Berkeley and Stanford. Agglutination-PCR detects autoantibodies in the solution-phase to ensure proper folding of antigen probes while leveraging the sensitivity and multiplex power of standard qPCR instruments. This project employs an innovative synthetic strategy to prepare and optimize a large panel of probes at low cost. While a traditional synthetic route would take up to 3 months, the proposed strategy could reduce the time down to one week. In addition, the probe library for autoantibody detection will be validated using banked serum/plasma from healthy patients or patients with systemic lupus erythematosus (SLE). This experiment will serve as a powerful proof-of-principle, as SLE displays many distinct autoantibodies that are challenging to detect with other methods. The analytical sensitivity and reproducibility of the proposed product will be compared to standard protein microarrays. The platform will be tested to ensure reproducibility and ease-of-use. The probe library and protocols will function as a minimum viable product and the prototype for applications in other diseases.