The goal of this project is to develop advanced ultrafiltration (UF) devices for sample preparation that are faster and easier to use than existing devices. New technology based on proprietary concepts enables ultrafiltration to be practiced at low pressure, and without the need of centrifuges or other complex apparatus, resulting in a single-pass, tangential-flow ultrafiltration method. The specific aims of the project are: 1. Design and construct prototypes of single sample tangential flow, hollow fiber, ultrafiltration devices. 2. Characterize the operating parameters in terms of the processing time and concentration factor as function of fiber geometry, device configuration and vacuum levels for protein solutions, using BSA or mixtures of BSA and peptides as a model. 3. Evaluate the performance of this device on an ultrafiltration application in life science research and compare this performance with published data on the performance of conventional devices. Existing UF devices have inherent limitations which result in slow filtration, high variability, and require the use of centrifuges which is cumbersome and not compatible with the robotic sample handling systems used in many laboratories and necessary for high-throughput screening. The UF filtration technology developed in this project combines novel fluidic and geometrical configurations that provide higher throughput and compatibility with robotic systems. The technology developed as a result of the proposed project is expected to remove barriers in current methods, facilitate the adoption of new diagnostic procedures, and reduce the cost of existing methods. This research will lead to the development of improved technology and new, lower cost devices for the purification and isolation of biological samples. Potential applications of the technology developed in this project include disposable devices for single and multiple sample processing, as well as for large-scale systems for biopharmaceutical manufacturing. The proposed research is expected to produce technology and devices which will provide significant benefits to public health. The research is particularly relevant to improved capabilities for the monitoring of cardiovascular disease states, and biomarkers in general, and the NHLBI's national efforts to prevent, detect and control hypertension. The technology from this project is expected to improve clinical diagnostic methods involving physiological fluids such as blood and urine, which need to be processed quickly in advance of a clinical testing protocol. The proposed research is expected to produce technology for significant benefits to public health. The research is particularly relevant to improved capabilities for the monitoring of cardiovascular disease states relative to national efforts to prevent, detect and control hypertension. The technology from this project is expected to improve clinical diagnostic methods involving physiological fluids such as blood and urine, which need to be processed quickly in advance of a clinical testing protocol.
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