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

这项小型企业创新研究（SBIR）项目的更广泛影响/商业潜力是开发一种工具，以加速发现新的自身抗体生物标志物，用于人类疾病的早期检测和个性化治疗。自身抗体生物标志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，现有自身抗体分析方法的可重复性、成本和敏感性有限，阻碍了发现新的生物标志物来改善这些疾病的管理。一个具有高灵敏度和鲁棒性的高成本效益的自身抗体分析平台可以揭示当前方法可能错过的自身抗体库的变化。这种工具将成为发现新的生物标记物的宝贵渠道，增强检测、治疗和理解许多疾病的能力。在每年74亿美元的生物标志物发现阵列市场中，拟议的工具作为服务或微阵列试剂盒供客户使用，满足了一些客户的必备需求，包括寻求新的疾病靶点的生物制药公司，改进临床试验和追求更好地了解人类生物学的学术团体。这个SBIR一期项目建议开发一种高度复用和敏感的自身抗体生物标志物分析工具，以加快生物标志物的发现。该方法采用了加州大学伯克利分校和斯坦福大学开发的一种新技术——凝集- pcr。凝集- pcr在溶液阶段检测自身抗体，以确保抗原探针的适当折叠，同时利用标准qPCR仪器的灵敏度和多重功能。该项目采用了一种创新的合成策略，以低成本制备和优化大型探针面板。传统的合成路线需要长达3个月的时间，而拟议的策略可以将时间缩短到一周。此外，用于自身抗体检测的探针文库将使用健康患者或系统性红斑狼疮（SLE）患者的血清/血浆进行验证。该实验将作为一个强有力的原理证明，因为SLE显示出许多不同的自身抗体，这些抗体很难用其他方法检测到。该产品的分析灵敏度和重现性将与标准蛋白微阵列进行比较。该平台将进行测试，以确保再现性和易用性。探针库和协议将作为最小可行产品和原型应用于其他疾病。

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.