Development of Fiber Optic Biosensors for Detection of Cancer Molecular Markers

描述(由申请人提供)： 目前，用于癌症检测和诊断、预测治疗效果或评估临床结果的低成本高通量技术有限。认识到这一挑战，NIH/NCI的癌症预防研究小额资助计划(PAR-08-055)寻求资助与“与临床前癌症检测相关的生物学分析技术、方法和临床技术的识别、开发和评估”相关的“试点和可行性研究”。针对这一点，在这一应用中，我们建议开发一种新型的低成本光纤生物传感器工具(DIP-Probe)来测量分子生物标记物，这些分子生物标记物可以用于癌症的早期检测/诊断、监测治疗反应或预测临床结果。该设备将依靠组合锥形光纤生物传感器(CTFOB)探头和半导体激光激励，并将根据夹心免疫分析原理发挥作用。分析物的量将以荧光信号的形式测量。荧光信号将由逝去波激发产生。此外，可以使用多个DIP探头来测量单个样品中的多个标记。此外，我们的初步研究结果表明，与其他目前可用的检测分析系统(包括蛋白质印迹和基于微珠的Luminex免疫分析)相比，该工具能够使用更少的试剂(减少10-100倍)和时间(不到1小时)检测样品中的少量分析物(灵敏度提高10倍)。我们建议的研究将有三个具体目标。具体目标#1将重点放在通过优化探头参数(锥角、锥长和探头半径)以及采用现有的夹心免疫分析技术来增强光纤探头的荧光信号。在具体目标#2中，我们将重点提高检测速度和灵敏度，将催化报告沉积(CARD)方法应用于光纤探针。该技术将被应用于检测和定量单个样本(例如血清样本)中的多个标志物。具体目标3将是优化这项技术的成本、周转时间和试剂使用，并将其与其他检测技术进行比较。这些研究的成功完成导致了一种低成本、便携式光纤工具的开发，该工具将用于对几种人类恶性肿瘤的早期检测、诊断、治疗效果、预后和监测。

DESCRIPTION (provided by applicant): Currently the low-cost high-throughput technologies for the detection and diagnosis of cancer, predict the efficacy of therapy or assess the clinical outcomes are limited. In recognition of this challenge, the cancer Prevention Research Small Grant Program (PAR-08-055) of the NIH/NCI seeks to fund "pilot and feasibility studies" related to "identification, development, and evaluation of biological analytic techniques, methodologies, and clinical technologies relevant to pre-clinical cancer detection. In direct response to this PAR, in this application we propose to develop a novel low cost fiber optic biosensor tool (dip-probe) to measure the molecular biomarkers which can be utilized for early detection/diagnosis of cancer, monitor the therapy response or predict the clinical outcomes. This device will rely on combination tapered fiber optic biosensor (CTFOB) probe and diode laser excitation, and will function on a sandwich immunoassay principle. The amount of the analyte will be measured in the form of a florescence signal. The fluorescence signal will be generated by evanescent wave excitation. Additionally, several dip-probes can be used to measure multiple markers in a single sample. Furthermore, our preliminary findings suggest that this tool is capable of detecting small amounts of analytes in samples (10 times higher sensitivity) utilizing reduced amounts of reagents (10-100 folds less) and time (less than an hour) when compared to other currently available detection assay systems including Western blot, and bead-based Luminex immunoassay. Our proposed studies will have three specific aims. Specific aim # 1 will focus on enhancement of fluorescence signal by optimization of probe parameters (tapered angle, taper length and probe radius), and adoption of existing sandwich immunoassay techniques to fiber optic probes. In specific aim # 2 we will focus on increasing the detection speed and sensitivity by adopting catalyzed reporter deposition (CARD) method to fiber optic probes. The technique will be applied to detect and quantify multiple markers in a single sample (e.g. serum sample). Specific aim # 3 will be to optimize the cost, turnaround time and reagent usage of this technique and compare it with other detections techniques. Successful completion of these studies result in the development of a low cost, portable fiber optic tool which will be utilized in early detection, diagnosis, therapy efficacy, prognosis, and surveillance of several human malignancies.