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测试方法，用于测量用于诊断测试的样本中特定蛋白质的浓度。近年来，蛋白质定量已经开始在医疗诊断、学术研究和制药开发中发挥越来越重要的作用，这些行业加起来代表着138亿美元的全球市场。在这些领域中的每一个领域，额外的步骤直接促成了推动向新方法过渡的前8个因素中的5个，包括：错误增加、测试时间更长、技术人员成本更高、吞吐量减少和复杂性增加。拟议的技术以一种广泛适用的方式解决了这些领域的每一个问题，方法是使用估计仅导致每次测试成本增加1%的材料，将步骤总数减少高达50%。这与医疗诊断行业尤其相关，在该行业，速度和准确性对患者的治疗和治疗成功有着巨大的影响。此外，测试复杂性的降低是促进便携式设备开发的关键因素，该设备具有潜在的新的国内应用以及在第三世界国家更多地获得医学测试的机会。该SBIR第一阶段项目建议开发一种广泛适用的基于标记的免疫分析平台，其步骤比传统的免疫分析更少。基于标记的免疫分析需要独立的抗原和检测抗体结合步骤，而单步无标记免疫分析提供的灵敏度明显较低，这在该领域是一个公认的问题。该项目旨在开发一种替代传统夹心免疫分析的方法，它使用分子内结合动力学来结合抗原和检测抗体结合步骤，从而允许在典型持续时间的单一孵育后进行基于标记的靶标检测。该方法最初将以链霉亲和素-生物素结合为模型系统进行研究，以优化复合体的形成，并在单一孵育后演示基于荧光的检测。然后，该平台将被应用于抗体介导的心肌梗死指标心肌肌钙蛋白I的检测，然后将与传统的夹心免疫分析进行比较。在这项研究的结论中，我们的目标是为简化的免疫分析平台提供概念验证数据，并有可能结合免疫分析步骤，在速度、技术人员时间、准确性、吞吐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.