NER: Reusable Active Nanostructured Capture Devices for Proteomics and Metabolomics

美国国家科学基金会-活性纳米结构和纳米系统（ANN） （NSF 06-595）纳米探索性研究（NER）提案编号：cbet -0707507首席研究员：Epps III， thomas隶属：特拉华大学提案标题：NER：蛋白质组学和代谢组学的可重复使用活性纳米结构捕获设备蛋白质组学和代谢组学的可重复使用活性纳米结构捕获装置代谢物和肽是生物体对环境变化反应的关键指标，其分析目前受到样品分离技术瓶颈的限制。为了解决这个问题，我们建议创建选择性可重复使用的纳米结构化学捕获装置，以隔离和浓缩特定的代谢物和肽，基于它们的化学功能。这种选择性捕获将通过结合嵌段共聚物的自然自组装和选择性化学来实现功能化纳米孔膜，以捕获含有-SH的小分子。含有-SH的肽和代谢物对于维持、感知和响应细胞还原氧化（氧化还原）状态至关重要，将从微生物培养物和海洋微生物群落中取样。虽然目前的提案仅限于-SH代谢物和肽作为概念证明，但最终目标是生产一种设备，该设备可以在空间定义的区域中捕获基于各种化学功能的分子，以促进下游分析所需的高度浓缩和纯化形式的一系列代谢物的收集。嵌段共聚物自组装成具有数十个有序相的周期性结构，包括几个多重连续网络。通过利用这些网络中存在的巨大内表面积和相互连接的扩散途径，我们可以设计具有定制运输和化学结合特性的材料，从而能够分析环境和临床标本中发现的小型或稀释样品。本研究计划有三个具体目标。首先是设计和生成纳米多孔聚合物膜模板。嵌段共聚物模板将专门为网络形成和选择性膜模板设计，具有单独的块以提供机械和环境稳定性，限制和化学功能。这些模板会在PI中合成吗？s实验室。π吗?S在共聚物网络结构方面的经验将为网络膜的形成提供重要的见解。二是用巯基选择性基团功能化纳米结构膜孔，以主动捕获环境代谢产物。选择性蚀刻和功能化方案将用于生成纳米多孔sh选择膜。第三个也是最后一个目标是使用模型系统和相关实验样本来测试主动捕获装置的性能。Co-PI吗?s在模型和环境微生物系统的蛋白质组学和-SH代谢分析方面的经验提供了用不同复杂性样品测试设备的能力。这将提供结构-功能关系，确定设备的性能，为后续设计提供信息。该项目具有重要的潜力，为复杂的海洋微生物群落中存在的-SH代谢产物和肽提供有价值的数据。这个高度跨学科的项目将训练研究生和本科生解决纳米技术中的关键科学和工程挑战。学生将探索化学、化学工程、材料科学、生物学和海洋研究的各个方面，将他们置于纳米技术的最前沿。具体的更广泛的影响和教育举措侧重于增加代表性不足的群体参与化学科学。其中包括：为特拉华州立大学和ACS少数民族学者项目的本科生提供多学科的夏季研究和指导机会。他在特拉华大学（University of Delaware）的实验室工作，并在特拉华大学开设了一门相关的、令人兴奋的纳米材料课程。此外，PI？作为项目小组委员会成员、前少数民族学者和导师，他继续参与ACS少数民族学者项目，这使他处于一个理想的位置，可以通过提议的努力对科学界产生重大影响。

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.