SBIR Phase I: A Novel Immunoassay Platform for Faster and More Accurate Protein Detection

这项小型企业创新研究（SBIR）提案的更广泛影响/商业潜力将是开发一种简化的检测方法，用于测量诊断检测样品中特定蛋白质的浓度。近年来，蛋白质定量开始在医学诊断、学术研究和药物开发中发挥越来越重要的作用，这些行业共同代表了138亿美元的全球市场。在这些领域中，额外的步骤直接导致了推动向新方法过渡的8大因素中的5个，包括：错误增加、测试时间延长、技术人员成本增加、吞吐量降低和复杂性增加。所提出的技术以广泛适用的方式解决了这些领域中的每一个，通过使用估计每次测试成本仅增加1%的材料，将步骤总数减少了50%。这与医疗诊断行业尤其相关，因为速度和准确性对患者治疗和治疗成功有着巨大的影响。此外，在测试复杂性的降低是一个关键因素，使便携式设备的发展潜力，允许新的国内应用，以及增加获得医疗testinginthird world countries.This SBIR第一阶段项目提出了开发一个广泛适用的标记为基础的免疫分析平台，步骤比传统的免疫分析。本领域中公认的问题是，基于标记的免疫测定需要独立的抗原和检测抗体结合步骤，并且单步无标记免疫测定提供显著较低的灵敏度。该拟议项目旨在开发一种替代传统夹心免疫测定的方法，该方法使用分子内结合动力学将抗原和检测抗体结合步骤联合收割机以一种允许在典型持续时间的单次孵育后进行基于标记的靶标检测的方式结合。该方法最初将使用链霉亲和素-生物素结合作为模型系统进行研究，以优化复合物的形成，并在单次孵育后证明基于荧光的检测。 该平台将应用于心肌梗死指标心肌肌钙蛋白I的抗体介导检测。然后将该系统与常规夹心免疫测定进行比较。在本研究结束时，目标是为简化的免疫分析平台提供概念验证数据，该平台具有联合收割机免疫分析步骤的潜力，以在速度、技术人员时间、准确度、通量和复杂性方面提供高达两倍的改进。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) proposal will be the development of a simplified testing method for measuring the concentration of specific proteins within a sample for diagnostic testing. In recent years, protein quantification has begun to play an increasingly important role in medical diagnostics, academic research, and pharmaceutical development, industries that together represent a $13.8 billion global market. In each of these areas, additional steps directly contribute to 5 of the top 8 factors driving the transition to new methods including: increased error, longer testing time, higher technician costs, decreased throughput, and added complexity. The proposed technology addresses each of these areas in a broadly applicable fashion by reducing the overall number of steps by up to 50% using materials that are estimated to contribute to a mere 1% increase in the cost of each test. This is particularly relevant to the medical diagnostic industry, where speed and accuracy have an immense effect on patient treatment and therapy success. Further, the reduction in test complexity represents a key factor enabling the development of portable devices with the potential to allow for new domestic applications as well as increased access to medical testing in third world countries.This SBIR Phase I project proposes to develop a broadly applicable label-based immunoassay platform with fewer steps than conventional immunoassays. It is a well-established problem in the field that label-based immunoassays require independent antigen and detection antibody binding steps, and that single step label-free immunoassays provide significantly lower sensitivity. The proposed project aims to develop an alternative to the conventional sandwich immunoassay, which uses intramolecular binding kinetics to combine antigen and detection antibody binding steps in a fashion which allows for label based target detection following a single incubation of typical duration. This method will be initially studied using streptavidin-biotin binding as a model system to optimize complex formation and demonstrate fluorescent-based detection following a single incubation. This platform will then be applied to antibody mediated detection of the myocardial infarction indicator cardiac troponin I. This system will then be evaluated in comparison with the conventional sandwich immunoassay. At the conclusion of this research, the goal is to provide proof-of-concept data for a simplified immunoassay platform with the potential to combine immunoassay steps to provide up to two-fold improvements in speed, technician time, accuracy, throughput, and complexity.