Sensitive, Integrating Multi-Waveguide Biosensor

描述（由申请人提供）：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，而其他患者则具有侵袭性临床结果，中位生存期显著降低。新的治疗方法可以诱导完全缓解，使前景不佳的患者在仍无症状的情况下接受治疗。然而，低肿瘤负荷的无症状患者（Binet A期）没有从治疗中获益，应该避免不必要的折磨和费用。问题是如何区分那些患有侵袭性疾病的患者和那些患有惰性疾病的患者。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细胞中不存在。Rosenwald等人在2001年首次发现了侵袭性CLL患者b细胞中存在ZAP-70的证据，Wiestner等人在2003年证实了这一点。存在于b细胞中的ZAP-70是一种能够准确区分这两组患者的生物标志物，因此是一种非常重要的预后指标。目前的测试方法要么没有广泛使用，要么没有充分标准化。在第一阶段，我们证明了集成波导生物传感器技术可以通过夹心免疫测定和荧光检测来检测缓冲液和各种基质中非常低浓度的蛋白质和细胞。检测迅速，最快20分钟，而且是定量的。在这个II期SBIR中，我们将应用生物传感器检测被诊断为CLL的患者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急需的信息，以便了解CLL疾病的分期，确定适当的治疗时机和选择，并确认治疗后的最小残留疾病。将开发针对CLL患者和对照者的血液标本的测定方法、仪器和测试盒。最常见的人类白血病是b细胞慢性淋巴细胞白血病。慢性淋巴细胞白血病缓慢进展亚型的患者可能有正常的寿命，可能永远不需要治疗。进展更迅速的CLL患者通常会发展为有症状的疾病，需要化疗。延迟化疗直至疾病进展的原则是建立在观察到在诊断时对所有患者进行化疗并非对所有患者都有益的基础上的。发现b细胞中的ZAP-70是疾病进展的可靠标志。该提案将开发一种对b细胞中ZAP-70的敏感检测方法，使医生能够识别侵袭性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.