NER: Reusable Active Nanostructured Capture Devices for Proteomics and Metabolomics

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National Science Foundation - Active Nanostructure and Nanosystems (ANN) (NSF 06-595)Nanoscale Exploratory Research (NER)Proposal Number: CBET-0707507Principal Investigator: Epps III, ThomasAffiliation: University of DelawareProposal Title: NER: Reusable Active Nanostructured Capture Devices for Proteomics and MetabolomicsNER: Reusable Active Nanostructured Capture Devices for Proteomics and MetabolomicsMetabolites and peptides are critical indicators of organism responses to environmental changes, whose analysis is currently limited by technological bottlenecks in sample fractionation. To address this problem, we propose to create selective reusable nanostructured chemical capture devices to sequester and concentrate specific metabolites and peptides, based on their chemical functionalities. This selective capture will be achieved by combining the natural self-assembly of block copolymers with selective chemistry to functionalize nanoporous membranes for the capture of -SH containing small molecules. -SH containing peptides and metabolites are critical for maintaining, sensing, and responding to cellular reductionoxidation (redox) status and will be sampled from microbial cultures and marine microbial communities. While the current proposal is limited to -SH bearing metabolites and peptides as a proof of concept, the ultimate goal is to produce a device that captures molecules based on a wide variety of chemical functionalities in spatially defined regions to facilitate the collection of an array of metabolites in the highly concentrated and purified forms required for downstream analyses.Block copolymers self-assemble into periodic structures characterized by dozens of ordered phases, including several multiply-continuous networks. By taking advantage of the immense internal surface area and interconnected diffusion pathways present in these networks, we can engineer materials with tailored transport and chemical binding properties, enabling the analysis of small or dilute samples such as those found in environmental and clinical specimens. There are three specific aims of this research proposal. The first is to design and generate nanoporous polymer membrane templates. Block copolymer templates will be specifically designed for network formation and selective membrane templating, with separate blocks to provide mechanical and environmental stability, confinement, and chemical functionality. These templates will be synthesized in the PI?s laboratory. The PI?s experience with copolymer network structures will provide significant insight into network membrane formation. The second is to functionalize nanostructured membranes pore with thiol selective moieties for the active capture of environmental metabolic products. Selective etching and functionalization protocols will be utilized to generate nanoporous -SH selective membranes. The third and final aim is to test the active capture device performance using both model systems and relevant experimental samples. The Co-PI?s experience in proteomic and -SH bearing metabolite analyses of model and environmental microbial systems provides the capability to test the device with samples of varied complexity. This will provide structure-function relationships, determining the performance of the device to inform subsequent designs. This project has significant potential to provide valuable data on -SH bearing metabolites and peptides present in complex marine microbial communities.This highly interdisciplinary project will train both graduate and undergraduate students to address key scientific and engineering challenges in nanotechnology. Students will explore aspects of chemistry, chemical engineering, materials science, biology, and marine studies, placing them at the forefront of nanotechnology. Specific broader impact and educational initiatives are focused on increasing the participation of underrepresented groups in the chemical sciences. These include: providing multidisciplinary summer research and mentorship opportunities for Delaware State University and ACS Minority Scholars Program undergraduate students in the PI and Co-PI?s labs at the University of Delaware (UD), and developing a relevant and stimulating nanomaterials course at the UD. Additionally, the PI?s continued involvement in the ACS Minority Scholars Program as a member of the Program subcommittee, former Minority Scholar, and mentor places him in an ideal position to have a substantial impact on the scientific community through the proposed endeavors.