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Protein/Peptide Separation with Polymer Brush Nanosponges

使用聚合物刷纳米海绵分离蛋白质/肽

基本信息

批准号：
7366938
负责人：
Daniel John Dyer
金额：
$ 21.41万
依托单位：
SOUTHERN ILLINOIS UNIVERSITY CARBONDALE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7366938
关键词：
Acrylates Adsorption Atomic Force Microscopy Biomedical Research Blood Bradykinin Charge Chemistry Complex Coupled Culture Media Cyanobacterium Doctor of Philosophy Dyes Escherichia coli Film Fingerprint Fluorescein Fluoresceins Fluorescence Fluorescence Spectroscopy Fluorescent Dyes Foundations Fractionation Funding Gel Chromatography Goals Gold Grant Health Human Individual Ions Laboratories Lasers Length Liquid substance MALDI-TOF Mass Spectrometry Mass Spectrum Analysis Measurement Measures Mentors Methacrylates Methodology Methods Modification Numbers Other Resources Oxides Pattern Peptides Plasma Pliability Polymers Porifera Property Proteins Proteome Proteomics Public Health Quartz Relative (related person)Research Sampling Schools Signal Transduction Silicon Solubility Solutions Solvents Source Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Spectrum Analysis Structure Students Surface Swelling Synthesis Chemistry Techniques Thick Time Universities Urine Vertebral column Work absorption acrylate aqueous base carboxylate copolymer density design detector feeding functional group hydrogel film infrared spectroscopy interest light scattering metabolomics monolayer monomer nanometer novel polymerization protonation research study response rhodamine 6G size uptake

项目摘要

DESCRIPTION (provided by applicant): We will examine the adsorption of peptides and fluorescent dye molecules onto nanometer length-tethered polymer films (so-called polymer brushes) in order to rapidly fractionate mixtures of compounds and analyze the components by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. These hydrogel films can swell by more than an order of magnitude in aqueous solutions, depending on the pH. More specifically, the anionic brushes expand at high pH (>6) and collapse at low pH(<6) due to protonation of the anionic carboxylates; we refer to these films as nanosponges since the collapse triggers a release of the solvent (i.e. the sponge is squeezed). And this release of solvent offers a method to deliver an analyte that had previously been fractionated by selective adsorption from a mixture. In this two-year project we are primarily interested in examining the loading capacity of the brush films (i.e. the amount of adsorbed peptide per unit volume), along with the various structural parameters that control the loading and release of compounds from the brush film. Furthermore, we will examine the temporal response of uptake and release of analytes, along with the collapse and expansion of the brush films. Since these are nanofilms we anticipate that their response will be faster than traditional bulk hydrogel films. The specific objectives of this work include the following: 1) Design and synthesize polymer brush films that will adsorb peptides and/or ionic dyes based on the charge distribution of the molecules and the brush; 2) Demonstrate the rapid uptake and release of the peptides or fluorescent dyes by altering the pH of the solution; 3) Develop structure-property relationships that allow us to maximize the loading capacity and the selectivity of the adsorbed species; 4) Evaluate the efficacy of this approach for peptide/protein fractionation as applied to MALDI analysis for proteomic and metabolomic applications; and 5) Introduce our respective graduate students to biomedical research and encourage undergraduates to pursue Ph.D. degrees in related fields. The films will be synthesized by "grafting from" methodology from self-assembled monolayers that are tethered to gold coated silicon wafers. The resulting films will be characterized by a variety of techniques including: RAIRS, ellipsometry, QCM, AFM, and time-resolved fluorescence spectroscopy. This project is relevant to public health since our long-term goal is to rapidly separate proteins and other biologically relevant molecules from various fluids such as blood, urine, or cultured media. These novel materials act like smart sponges to selectively adsorb specific peptides based on ionic interactions, which will then be analyzed by mass spectrometry.

描述（由申请人提供）：我们将检查肽和荧光染料分子在纳米长度系留聚合物膜（所谓的聚合物刷）上的吸附，以快速破碎化合物的混合物，并通过基质辅助激光解吸电离飞行时间（MALDI-TOF）质谱法分析组分。这些水凝胶膜可以在水溶液中溶胀超过一个数量级，这取决于pH。更具体地说，由于阴离子羧酸盐的质子化，阴离子刷在高pH（>6）下膨胀，在低pH（<6）下塌陷;我们将这些膜称为纳米海绵，因为塌陷触发溶剂的释放（即海绵被挤压）。这种溶剂的释放提供了一种方法来传递先前通过选择性吸附从混合物中分离的分析物。在这个为期两年的项目中，我们主要感兴趣的是检查刷膜的负载能力（即每单位体积的吸附肽的量），沿着的各种结构参数，控制加载和释放的化合物从刷膜。此外，我们将检查的时间响应的吸收和释放的分析物，随着沿着的崩溃和扩张的刷膜。由于这些是纳米膜，我们预计它们的响应将比传统的块状水凝胶膜更快。本论文的具体目标包括：1）根据分子和刷的电荷分布，设计并合成能够吸附肽和/或离子染料的聚合物刷膜; 2）通过改变溶液的pH值，证明肽或荧光染料的快速吸收和释放; 3）开发结构-性质关系，使我们能够最大化吸附物种的负载能力和选择性; 4）评估这种用于肽/蛋白质分级分离的方法在应用于蛋白质组学和代谢组学应用的MALDI分析时的功效;（五）介绍研究生从事生物医学研究，鼓励本科生攻读博士学位。相关领域的学位。这些薄膜将通过“嫁接”方法从拴在镀金硅片上的自组装单层合成。所得薄膜将通过多种技术表征，包括：RAIRS、椭圆偏振法、QCM、AFM和时间分辨荧光光谱。该项目与公共卫生有关，因为我们的长期目标是从血液，尿液或培养基等各种液体中快速分离蛋白质和其他生物相关分子。这些新型材料就像智能海绵一样，基于离子相互作用选择性地吸附特定的肽，然后通过质谱法进行分析。