Innovative Tools for the Molecular Diagnosis of Cancer

描述（由申请人提供）：美国国家癌症研究所呼吁为癌症研究和诊断提供准确、实用和负担得起的新工具。Alderon Biosciences, Inc.提议通过开发一种开放的生物传感器平台技术来满足这一需求，用于癌症基因表达的定量研究，目标是取代速度较慢且成本较高的方法。本次SBIR提案“Innovative Tools for The Molecular Diagnosis of Cancer”的工作假设是Alderon创新的用于癌症基因表达分析的酶增强电化学系统（essystem）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量化了差异基因表达。第一阶段工作取得的主要里程碑是证明了目标检测灵敏度足以用于酶增强探针检测小组织样本中甚至很少表达的信息，而无需目标扩增（在小到104个细胞的样本中，将mRNA目标量化到每个细胞1-3个拷贝）。在第二阶段，将进一步开发和验证具有小传感器面积电极的新型eSystem读取器和ePlates，这些电极需要非常少量的样品和试剂。II期项目目标是：1)制作和测试新的系统组件（改进的检测方法，小传感器ePlates和ePlate阅读器），用于同时定量多个癌症基因靶点的表达水平，而无需使用靶点扩增步骤；2)生产新的essystem读取器和96传感器微孔板的制造原型；3)改进性能，直到与定量实时RT-PCR分析相比，使用该系统获得等同或更好的基因表达分析（速度、样本量、成本、可靠性）。高灵敏度的基因表达测量，没有现有方法的复杂性和成本，已经被证明是可能的，新的小传感器面积微孔板，需要非常小体积的样品和试剂。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5-7年内进入临床使用时，每种主要疾病的市场可能会增长到每年超过10亿美元。我们打算借助成熟的技术和战略合作伙伴关系，在研究和测试方面取得战略份额。

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.