Sensitive, Integrating Multi-Waveguide Biosensor

描述(申请人提供)：慢性淋巴细胞白血病(CLL)是一种异质性疾病。许多患者从不需要治疗，而另一些患者的临床结果积极，中位生存期显著降低。可以导致完全缓解的新疗法允许前景不佳的患者在仍然没有症状的情况下接受治疗。然而，无症状的肿瘤负担低的患者(比奈A期)从治疗中得不到任何好处，应该免除不必要的磨难和费用。问题是如何区分哪些患者患有侵袭性疾病，哪些患者适合接受治疗，而哪些患者不是慢性病患者。ZAP-70是一种细胞内蛋白，在T细胞和自然杀伤细胞中表达，但在健康人的B细胞中不表达。Rosenwald等人首次证实侵袭性CLL患者的B细胞中存在ZAP-70。2001年，并由Wiestner等人核实。2003年。B细胞中存在的ZAP-70是一种生物标记物，可以准确区分这两个患者组，因此是一个非常重要的预后指标。目前的测试方法要么不是广泛可用的，要么是不够标准化的。在第一阶段，我们展示了集成波导生物传感器技术可以通过夹心免疫分析和荧光检测来检测缓冲液和各种基质中极低浓度的蛋白质和细胞。检测迅速，最快可达20分钟，而且是定量的。在这个第二阶段的SBIR中，我们将应用该生物传感器检测已被诊断为CLL的患者的B细胞中的ZAP-70。检测到的ZAP-70水平将为医生和患者提供有用和抢手的信息，以便了解CLL疾病的阶段，确定适当的治疗时机和选择，并确认治疗后最小的残留病。将开发分析、仪器和测试试剂盒，并针对CLL患者和对照组的血液样本进行测试。人类最常见的白血病是B细胞慢性淋巴细胞白血病(CLL)。慢性淋巴细胞性白血病进展缓慢的患者可能有正常的寿命，可能永远不需要治疗。进展较快的慢性淋巴细胞性白血病患者通常进展为有症状的疾病，需要化疗。推迟化疗直到病情进展的原则是建立在观察到所有患者在确诊时给予化疗并不是对所有人都有利的基础上的。B细胞中的ZAP-70被发现是疾病进展的可靠标志。这项提议将开发一种B细胞中ZAP-70的灵敏检测方法，使医生能够识别具有侵袭性形式的CLL患者，并立即提供治疗。

DESCRIPTION (provided by applicant): Chronic lymphocytic leukemia (CLL) is a heterogeneous disease. Many patients never need treatment, whereas others have an aggressive clinical outcome with significantly reduced median survival. New treatments that can induce complete remission permit patients with poor outlook to be treated while they are still asymptomatic. However, asymptomatic patients with low tumor burden (Binet stage A) derive no benefit from treatment and should be spared the unnecessary ordeal and expense. The problem is how to distinguish those patients with aggressive disease and who are candidates for treatment from those with indolent disease who are not. ZAP-70 is an intracellular protein that is expressed in T-cells and natural killer cells, but absent in B-cells of healthy people. First evidence that presence of ZAP-70 in B-cells of patients with aggressive form of CLL was identified by Rosenwald et al. in 2001 and verified by Wiestner et al. in 2003. ZAP-70 present in B-cells is a biomarker that can accurately differentiate these two patient groups and, thus, a very important prognostic indicator. Current test methods are either not widely available or insufficiently standardized. In Phase I, we demonstrated that the Integrating Waveguide Biosensor technology can be used to detect very low concentrations of proteins and cells in buffer and various matrices by sandwich immunoassay and fluorescence detection. Detection is rapid, as fast as 20 minutes, and quantitative. In this Phase II SBIR, we will apply the biosensor to detection of ZAP-70 in B cells of patients who have been diagnosed with CLL. The level of ZAP-70 so detected will provide useful and sought-after information for physicians and patients in order to understand the stage of CLL disease and determine the appropriate timing and choice of treatment, and to confirm post-treatment minimal residual disease. Assays, instrument, and a test cartridge will be developed and tested against blood specimens from CLL patients and controls. The most common human leukemia is B-cell chronic lymphocytic leukemia (CLL). Patients with a slowly progressive subtype of CLL may have a normal life span and may never require treatment. Patients with the more rapidly progressive form of CLL typically progress to symptomatic disease and need chemotherapy. The principle of delaying chemotherapy until the disease has progressed is founded on the observation that chemotherapy given to all patients at diagnosis is not beneficial to all. ZAP-70 in B-cells was found to be a reliable marker of disease progression. This proposal will develop a sensitive detection for ZAP-70 in B-cells to allow the doctor to identify the patients with aggressive form of CLL and provide treatment immediately.