Innovative Tools for the Molecular Diagnosis of Cancer

描述（由申请人提供）：国家癌症研究所呼吁癌症研究和诊断的新工具是准确的，实用的和负担得起的。Alderon Biosciences，Inc.提出通过开发用于癌症基因表达的定量研究的开放式生物传感器平台技术来满足这一需求，其目标是取代较慢且更昂贵的方法。SBIR关于“癌症分子诊断的创新工具”的提案的工作假设是，Alderon用于癌症基因表达分析的创新酶增强电化学系统（eSystem）的一次性传感器阵列和电化学读取器将极大地促进癌症生物标志物的研究。使用Alderon的生物传感器平台技术进行定量癌症基因表达分析的可行性已在I期工作中得到证实，并在诱导和非诱导癌细胞中定量了差异基因表达。在I期工作中实现的主要里程碑是证明了靶检测灵敏度足以在小组织样品中基于酶增强的探针检测甚至很少表达的信息，而不需要靶扩增（在小至104个细胞的样品中，mRNA靶定量低至每个细胞1-3个拷贝）。在第二阶段，将进一步开发和验证新的eSystem读取器和具有小传感器面积电极的ePlate，这些电极需要非常少量的样本和试剂。第二阶段项目的目标是：1）制作和测试新的eSystem组件（改进的测定、小传感器ePlate和ePlate读数器），用于同时定量多个癌症基因靶标的表达水平，而不使用靶标扩增步骤; 2）生产新的eSystem读数器和96传感器微孔板的制造原型;和3）改进性能，直到与定量实时RT-PCR测定相比，用eSystem获得等同或更好（速度、样品大小、成本、可靠性）的基因表达测定。高灵敏度的基因表达测量没有现有方法的复杂性或成本已被证明是可能的新的小传感器面积的微孔板，需要非常小的样品和试剂的体积。这种先进的电化学技术将进一步发展用于癌症研究和诊断，然后扩展到其他人类健康产品。当疾病相关的基因表达产品在估计的5-7年内达到临床用途时，每种主要疾病的市场可能会增长到超过10亿美元/年。我们打算在成熟的技术和战略合作伙伴关系的帮助下，在研究和测试领域占据战略市场份额。

DESCRIPTION (provided by applicant): The National Cancer Institute has called for new tools for cancer research and diagnosis that are accurate, practical and affordable. Alderon Biosciences, Inc. proposes to fill this need by developing an open biosensor platform technology for quantitative studies of cancer gene expression, with the goal of replacing slower and more costly methods. The working hypothesis of this SBIR proposal on "Innovative Tools for the Molecular Diagnosis of Cancer'' is that the disposable sensor arrays and electrochemical readers of Alderon's innovative enzyme-enhanced electrochemical system (eSystem) for cancer gene expression analysis will greatly facilitate studies of cancer biomarkers. Feasibility of using Alderon's biosensor platform technology for quantitative cancer gene expression analysis has been demonstrated in Phase I work and differential gene expression was quantified in induced and non-induced cancer cells. The major milestone achieved in Phase I work was demonstration of target detection sensitivity sufficient for enzyme-enhanced probe-based detection of even rarely expressed messages in small tissue samples without need for target amplification (quantification of mRNA targets down to 1-3 copies per cell in samples as small as 104 cells). In Phase II, new eSystem readers and ePlates with small sensor-area electrodes that require very small volumes of samples and reagents will be further developed and validated. Phase II Project Aims are: 1) make and test new eSystem components (improved assays, small-sensor ePlates and ePlate readers) for simultaneous quantification of levels of expression of multiple cancer gene targets without use of target amplification steps; 2) produce manufacturing prototypes of new eSystem readers and 96-sensor microplates; and 3) refine performance until equivalent or better (speed, sample size, cost, reliability) assays of gene expression are obtained with the eSystem compared to quantitative real-time RT-PCR assays. Highly sensitive gene expression measurements without the complexity or cost of existing methods have been proven possible with new small sensor-area microplates that require very small volumes of samples and reagents. This advanced electrochemical technology will be further developed for cancer research and diagnostics, and then extended to other human health products. When disease-related gene expression products reach clinical use in an estimated 5-7 years, markets will likely grow to exceed a billion dollars/yr for each major disease. We intend to reach a strategic share of the market in research and testing with the aid of proven technology and strategic partnerships.