Novel Polymer Adsorbents for Selective Peptide and Small Protein Separations from Blood

用于从血液中选择性分离肽和小蛋白的新型聚合物吸附剂

• 批准号: 1604170
• 负责人: Roberto Guzman
• 金额: $ 31.77万
• 依托单位: University of Arizona
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1604170&HistoricalAwards=false
• 关键词: Novel; Polymer; Adsorbents; Selective; Peptide

Proposal Number: 1604170, GuzmanNovel Adsorbents for Selective Peptide and Small Protein Separations from Blood Small biomolecules or biomarkers from human blood or other biological fluids such as urine are usually found in trace amounts. Their identification and purification is in general hindered by the high concentration of human albumin and other proteins found in blood and biological fluids. The effective recovery and identification of these biomolecules from complex biological mixtures has been until now a considerable challenge. The methods developed in this research will allow the effective isolation of bioactive peptides, hormones, and biomarkers that can be used in biomedical areas. This research will enable medical applications that isolate and remove toxins from biological fluids and then identify biomarkers for early cancer detection. In this research, selective adsorbents with polymers that do not allow the adsorption of large proteins will be synthesized. The adsorbents will be modified with chemical groups that will permit only the selective adsorption, from complex mixtures of targeted bioactive peptides, hormones and biomarkers that are found only at low concentrations. The technology will be tested in samples from biological fluids such as blood, urine and saliva. Studies will be conducted to obtain higher purification rates for targeted biomolecules. This will translate into lower operation costs in the development and discovery of new biomarkers and in the detoxification and cleaning of human blood. This project will be interdisciplinary in nature, the engineering, biology, and chemical knowledge used and gained in this research will be of interest to many disciplines. This program will involve undergraduate and graduate students from different disciplines with interest in chemistry, molecular biology, medicine, material sciences and chemical engineering.Despite many efforts to develop efficient protein purification techniques, the isolation of peptides and small proteins on a larger than analytical scale remains a significant challenge. There is currently no similar separation product commercially available. The primary objective of this research will be the development and characterization of a chromatographic adsorbent that allows separation of peptides and biomolecules by simultaneous specific adsorption, and size exclusion using polysaccharide matrices modified with permeation control polymers. This will be accomplished by grafting in chromatographic matrices two different heterogeneous ligands, one, a polymer such as polyethylene glycol that permits the permeation of only certain molecular size range compounds, and a second one, an affinity ligand such as a chelating agent or an ion exchange group that preferentially binds target biomolecules of that specific molecular size range that could cross the restricted polymer barrier. This method will eliminate competition for adsorptive sites on the matrix surface, between large and small molecules, thus increasing adsorption rate and capacity and effective separations and purifications of desired biomolecules. To effectively characterize the properties of these new matrices, systematic dual surface synthesis functionalization protocols will be developed using polyethylene glycol of different sizes and densities as the permeating polymer and chelating and ion exchange groups as affinity ligands.

提案编号：1604170，GuzmanNovel Adsorbents for Selective Peptide and Small Protein Separations from Blood来自人血液或其它生物流体如尿的小生物分子或生物标记物通常以痕量存在。它们的鉴定和纯化通常受到血液和生物流体中发现的高浓度人白蛋白和其他蛋白质的阻碍。从复杂的生物混合物中有效地回收和鉴定这些生物分子一直是一个相当大的挑战。在这项研究中开发的方法将允许有效分离生物活性肽，激素和生物标志物，可用于生物医学领域。这项研究将使医学应用能够从生物液体中分离和去除毒素，然后识别早期癌症检测的生物标志物。在这项研究中，将合成具有不允许吸附大蛋白质的聚合物的选择性吸附剂。吸附剂将用化学基团进行修饰，这些化学基团仅允许从仅在低浓度下发现的靶向生物活性肽、激素和生物标志物的复杂混合物中选择性吸附。该技术将在血液、尿液和唾液等生物液体样本中进行测试。将进行研究以获得目标生物分子的更高纯化率。这将转化为开发和发现新生物标志物以及人类血液解毒和清洁的较低运营成本。该项目本质上是跨学科的，本研究中使用和获得的工程、生物学和化学知识将引起许多学科的兴趣。该项目将涉及来自不同学科的本科生和研究生，他们对化学、分子生物学、医学、材料科学和化学工程感兴趣。尽管许多人努力开发高效的蛋白质纯化技术，但在分析规模以上分离肽和小蛋白质仍然是一个重大挑战。目前没有类似的分离产品可商购。本研究的主要目的将是开发和表征的色谱吸附剂，允许分离的肽和生物分子的同时特异性吸附，和尺寸排阻使用多糖基质改性的渗透控制聚合物。这将通过在色谱基质中接枝两种不同的非均相配体来实现，一种是聚合物如聚乙二醇，其仅允许某些分子大小范围的化合物渗透，第二种是亲和配体如螯合剂或离子交换基团，其优先结合可穿过受限的聚合物屏障的特定分子大小范围的靶生物分子。该方法将消除大分子和小分子之间对基质表面上的吸附位点的竞争，从而提高吸附速率和容量以及所需生物分子的有效分离和纯化。为了有效地表征这些新基质的性质，将使用不同尺寸和密度的聚乙二醇作为渗透聚合物，并使用螯合和离子交换基团作为亲和配体开发系统的双表面合成功能化方案。