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

这个小型企业创新研究(SBIR)项目的更广泛的影响/商业潜力将是开发一种平台技术，用于检测研究和临床诊断的自身抗体标志物。精确医学要求开发更强大的生物分析技术来诊断疾病和指导靶向治疗。该平台使用基于连接的DNA条形码技术来改进抗体检测，提高了分析灵敏度和多重能力，以在最临床有用的时间检测更多抗体。此外，作为一种溶液相检测，它能够检测出许多与临床相关的自身抗体，而这些抗体是常规技术(如ELISA)无法检测的。这一平台有可能加速人类各种疾病的救生诊断技术的发展。这个小企业创新研究第二阶段项目旨在开发第一个解决方案阶段、超灵敏和多重抗体分析平台，用于人类疾病的早期检测、监测和治疗。利用凝集-聚合酶链式反应(ADAP)平台进行抗体检测是多重免疫检测技术的重大进展。许多目前的多重技术，如微阵列和基于珠基的阵列，由于交叉分析物干扰而规模化程度较差，并且缺乏在最有利的时间检测致残性疾病的分析灵敏度。第一阶段的结果表明，扩展和验证的ADAP技术用于抗体生物标记物的超灵敏和多重检测。第二阶段的目标是：1)验证多重ADAP分析用于临床诊断的有效性，为自身免疫性结缔组织疾病、甲状腺疾病和乳糜泻疾病服务；2)将多重ADAP分析技术高通量自动化用于临床诊断；3)制造分析试剂和建立质量控制标准；以及4)证明多重ADAP对更广泛的抗体的适用性。该项目的智力价值在于实现了10,000倍的灵敏度提高，20-30%的多路传输特异度增加，同时可以选择使用现有的PCR阅读器平台来帮助保持较低的实验室资本成本。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be to develop a platform technology for detecting autoantibody markers for research and clinical diagnostics. Precision medicine requires the development of more powerful bioanalytic technologies to diagnose disease and direct targeted therapies. This platform uses a ligation-based DNA barcoding technology for improved antibody detection with increased analytical sensitivity and multiplex power to detect more at the most clinically useful time. Additionally, as a solution-phase assay, it is able to detect numerous clinically-relevant autoantibodies that are refractory to common techniques like ELISA. This platform has the potential to accelerate the development of lifesaving diagnostics across a broad spectrum of human diseases. This Small Business Innovation Research Phase II project aims to develop the first solution-phase, ultrasensitive and multiplex antibody assay platform for the early detection, monitoring and treatment of human diseases. The Antibody Detection by Agglutination-PCR (ADAP) platform represents a major advancement in multiplex immunoassay testing. Many current multiplex technologies, such as microarrays and bead-based arrays, scale poorly due to cross-analyte interference, and lack the analytical sensitivity to detect crippling diseases at the most favorable time. The Phase I results showed the expanded and validated ADAP technology for the ultrasensitive and multiplex detection of antibody biomarkers. The Phase II objectives are: 1) Validation of the multiplex ADAP assay for clinical diagnostics serving autoimmune connective tissue diseases, thyroid disorders and celiac diseases; 2) High-throughput automation of multiplex ADAP assay technology for clinical diagnostics; 3) Manufacturing of assay reagents and establishment of quality control standards; and 4) Demonstration of the applicability of multiplex ADAP for a broader set of antibodies. The intellectual merit of this project resides in the achievement of 10,000x increased sensitivity, and 20-30% increased specificity at multiplex while having the option to use existing PCR reader platforms to help keep lab capital costs low.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.