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，而另一些患者则具有积极的临床结果，中位生存期显着降低。可以诱导完全缓解的新疗法允许前景不佳的患者在仍然无症状时接受治疗。然而，低肿瘤负荷（Binet分期A）的无症状患者不能从治疗中获益，应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不适合治疗的惰性疾病患者。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细胞中不存在。ZAP-70存在于具有侵袭性形式的CLL患者的B细胞中的第一证据由Rosenwald等人在2001年鉴定，并由Wiestner等人在2003年验证。存在于B细胞中的ZAP-70是一种生物标志物，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用于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。检测快速，最快20分钟，定量。在这个第二阶段SBIR中，我们将应用生物传感器检测已被诊断为CLL的患者的B细胞中的ZAP-70。如此检测的ZAP-70水平将为医生和患者提供有用和受欢迎的信息，以了解CLL疾病的阶段，并确定适当的治疗时机和选择，并确认治疗后的微小残留疾病。将开发检测试剂盒、仪器和检测试剂盒，并针对CLL患者和对照的血液标本进行检测。最常见的人类白血病是B细胞慢性淋巴细胞白血病（CLL）。慢性淋巴细胞白血病的缓慢进展亚型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白血病患者通常会发展为有症状的疾病并需要化疗。延迟化疗直到疾病进展的原则是基于这样的观察，即在诊断时给予所有患者化疗并不是对所有人都有益。发现B细胞中的ZAP-70是疾病进展的可靠标志物。该提案将开发一种灵敏的检测B细胞中ZAP-70的方法，使医生能够识别患有侵袭性CLL的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.