Multiplexed Assays on the BioCD for Acute Lymphocytic Leukemia

描述（由申请人提供）：BioCD是一种新兴的无标记检测技术，具有高复用和高通量的潜力，可同时在许多样品中筛选许多分析物。BioCD技术的基础是快速光学干涉扫描。干涉测量法是直接光学检测的最灵敏和最定量的手段。它比荧光更快，具有更好的信噪比，并且不需要标记，这对于多分析物检测至关重要。该提案的广泛和长期目标是首次将BioCD应用于癌症的预后。其目标是在300名患者的大型队列中检测多种生物标志物，以以前免疫组织学阵列无法达到的水平，预测患者对化疗的反应结果。该提案的具体目的是建立一组与急性淋巴细胞白血病（ALL）患者化疗结果预测相关的生物标志物的标准反应曲线，并在BioCD上进行的测定与之前在有限数量的组织微阵列上进行的测定之间进行比较研究。接下来的目标是扩大BioCD的能力，以筛选已经从300名急性淋巴细胞白血病患者队列中收集的一组样本中的扩展生物标志物集。研究设计和实现所述目标的方法依赖于高容量的自动蛋白质点样仪和高速激光干涉读取器。将使用组织裂解物中的市售抗原建立标准浓度曲线，然后测量健康组织裂解物中的标志物浓度。这些测量值设定了金标准，将与大型患者队列中测量的扩展标记物集进行比较。这项研究与公共卫生的相关性是标记物和样本基础的扩展，以建立标准风险急性淋巴细胞白血病患者分子途径失活的预后价值的更强的临床确认，并建立BioCD作为一种新的高容量资源用于癌症的诊断和预后应用。

DESCRIPTION (provided by applicant): The BioCD is an emerging label-free assay technology with potential for high multiplexing and high throughput to screen for many analytes across many samples simultaneously. The basis of the BioCD technology is rapid optical interferometric scanning. Interferometry is the most sensitive and quantitative means of direct optical detection. It is faster than fluorescence, with better signal-to-noise, and it requires no labels, which is essential for multiple analyte detection. The broad, long-term objectives of this proposal are to apply the BioCD for the first time to the prognosis of cancer. The goal is to assay multiple biomarkers across a large cohort of 300 patients, at a level previously inaccessible to immunohistological arrays, to predict patient outcome in response to chemotherapy. The specific aims of this proposal are to establish standard response curves for a set of biomarkers relevant for predicting chemotherapy outcome for patients with acute lymphocytic leukemia (ALL), and to do a comparative study between assays performed on the BioCD and assays performed previously on a limited number of tissue microarrays. This is followed by the aim to scale up the capacity of the BioCD to screen for an expanded biomarker set across a set of samples already collected from the cohort of 300 patients with acute lymphocytic leukemia. The research design and the methods for achieving the stated goals rely on high- capacity automated protein spotters and high-speed laser interferometric readers. Standard concentration curves will be established using commercially available antigens in tissue lysates, followed by measurements of marker concentrations in healthy tissue lysate. These measurements set the gold standard against which the expanded marker set measured across the large patient cohort will be compared. The relevance of this research to public health is the expansion of the marker and sample base to establish stronger clinical confirmation of the prognostic value of inactivation of a molecular pathway in patients with standard-risk acute lymphocytic leukemia, and to establish the BioCD as a novel high-capacity resource for diagnostic and prognostic applications for cancer.