Phi29 motor nanopore for single molecule sensing

 描述(申请人提供)：乳腺癌通常在疾病的早期阶段没有症状。目前最先进的筛查工具，如乳房X光摄影和乳房检查，由于没有肿块病变而漏掉了早期乳腺癌，对年轻女性的癌症检测无效，需要侵入性活组织检查。需要新的方法来补充现有的乳腺癌前病变和癌前病变的检测和预后方法，因为这些方法很小，更容易治疗。分析乳头抽吸液中分泌的糖抗原、蛋白抗原和非编码microRNAs(MiRNA)是一种非常有前途的方法，能够直接从反映乳腺组织特异性肿瘤微环境的体液中检测与癌症相关的生物标记物。我们的长期目标是开发一个简单、灵敏和强大的检测平台，以便在无症状的早期阶段诊断疾病。我们的诊断平台基于优雅而精细的生物马达蛋白质通道。我们将马达通道插入到脂质双层中，当分析物与通道相互作用时，得到的系统显示出坚固的特性，并产生极其敏感、稳定和可重现的电导信号。我们进一步证明了它作为一种检测癌症相关蛋白的高灵敏设备的有效性。这项提议的短期目标是开发这种运动通道作为纳米孔，用于高通量检测从乳腺癌患者乳头抽吸液中提取的糖抗原、蛋白质抗原和miRNA生物标志物的单分子。该平台包括：作为高灵敏纳米孔的膜嵌入电机通道；用于捕获乳腺癌相关抗原的可编程探针；以及用于多路高通量单分子检测的超灵敏电生理装置。将我们的设备融入常规实践中，将使医生能够更早地监测乳腺导管内的细胞变化，以免它们发展为更大的、潜在的癌症病变。

 DESCRIPTION (provided by applicant): Breast cancer is generally asymptomatic at early stages of the disease. Current state of the art screening tools such as mammography and breast examination miss early breast cancers due to the absence of mass lesions, are ineffective in detecting cancer in young women, and require invasive biopsy. New methods are needed to complement current methods for the detection and prognosis of precancerous and cancerous breast lesions while they are small and more easily treated. The analysis of the secreted glycoantigens, protein antigens and non-coding microRNAs (miRNA) in nipple aspirate fluid represents a very promising approach able to detect cancer-related biomarkers directly in body fluids mirroring the breast tissue-specific tumor microenvironment. Our long term goal is to develop a simple, sensitive and robust detection platform for diagnosing diseases at an asymptomatic early stage. Our diagnostic platform is based on the elegant and elaborate protein channel of a biological motor. We have inserted the motor channel into a lipid bilayer and the resulting system exhibits robust properties and generates extremely sensitive, stable and reproducible conductance signatures when analytes interact with the channel. We further demonstrated its utility as a highly sensitive device for detection of cancer-associated proteins i presence of many contaminants at single molecule level. The short-term goal of this proposal is to develop this motor channel as a nanopore for high-throughput single molecule detection of glyco-antigen, protein antigen and miRNA biomarkers extracted from breast cancer patient nipple aspirate fluid. The platform consists of: a membrane-embedded motor channel serving as a highly sensitive nanopore; programmable probes for capturing breast cancer associated antigens; and ultra-sensitive electrophysiological setup for multiplexed high-throughput single molecule detection. Incorporating our device into routine practice will enable physicians to monitor cellular changes within breast ducts earlier, before they develop into larger, potentially cancerous lesions.