Phi29 motor nanopore for single molecule sensing

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 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.