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。精准医疗需要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，提高分析灵敏度和多重能力，以便在临床上最有用的时间检测更多。此外，作为一种溶液相测定法，它能够检测许多临床相关的自身抗体，这些抗体是ELISA等常用技术难以检测的。 该平台有可能加速开发针对各种人类疾病的救命诊断方法。该小型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，超灵敏和多重抗体检测平台，用于早期检测，监测和治疗人类疾病。抗体检测-聚合酶链反应（ADAP）平台代表了多重免疫分析检测的重大进步。许多当前的多重技术，如微阵列和基于珠的阵列，由于交叉分析物干扰而规模很小，并且缺乏在最有利的时间检测致残性疾病的分析灵敏度。 第一阶段的结果显示了扩展和验证的ADAP技术，用于抗体生物标志物的超灵敏和多重检测。第二阶段的目标是：1）用于自身免疫性结缔组织疾病、甲状腺疾病和乳糜泻的临床诊断的多重ADAP测定的验证; 2）用于临床诊断的多重ADAP测定技术的高通量自动化; 3）测定试剂的制造和质量控制标准的建立;以及4）证明多重ADAP对更广泛的抗体组的适用性。该项目的智力价值在于实现了10，000倍的灵敏度提高，以及20-30%的多重特异性提高，同时可以选择使用现有的PCR读取器平台，以帮助保持实验室资本成本低。该奖项反映了NSF的法定使命，并被认为值得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来支持。

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.