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

建议编号：1604170，Guzman Novel吸着剂用于从血液中分离选择性多肽和小蛋白人体血液或其他生物液体(如尿液)中的小生物分子或生物标志物通常存在于微量中。它们的鉴定和纯化通常受到血液和生物液中发现的高浓度人类白蛋白和其他蛋白质的阻碍。到目前为止，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。这项研究开发的方法将允许有效地分离生物活性多肽、激素和可用于生物医学领域的生物标记物。这项研究将使从生物液中分离和去除毒素，然后识别用于早期癌症检测的生物标记物的医学应用成为可能。在这项研究中，将合成带有不允许吸附大分子蛋白质的聚合物的选择性吸附剂。吸附剂将被化学基团修饰，仅允许从仅在低浓度下发现的目标生物活性多肽、激素和生物标记物的复杂混合物中进行选择性吸附。这项技术将在血液、尿液和唾液等生物液体样本中进行测试。将进行研究，以获得更高的目标生物分子提纯率。这将转化为在开发和发现新的生物标记物以及人类血液的解毒和清洁方面降低运营成本。这个项目将是跨学科的，在这项研究中使用和获得的工程学、生物学和化学知识将对许多学科产生兴趣。这个项目将包括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。尽管在开发高效的蛋白质纯化技术方面做出了许多努力，但在比分析规模更大的范围内分离多肽和小蛋白质仍然是一个巨大的挑战。目前市场上还没有类似的分离产品。这项研究的主要目标是开发和表征一种色谱吸附剂，这种吸附剂可以通过同时进行特定吸附和使用渗透控制聚合物修饰的多糖基质来排除尺寸，从而实现多肽和生物分子的分离。这将通过在层析基质中嫁接两种不同的非均相配体来实现，一种是仅允许某些分子尺寸范围的化合物渗透的聚合物，如聚乙二醇，另一种是亲和配体，如螯合剂或离子交换基团，其优先结合可跨越受限聚合物屏障的特定分子尺寸范围的目标生物分子。这种方法将消除大小分子之间对基质表面吸附位置的竞争，从而提高吸附速度和容量，并有效地分离和纯化所需的生物分子。为了有效地表征这些新基质的性质，将以不同大小和密度的聚乙二醇为渗透聚合物，以螯合和离子交换基团为亲和配体，开发系统的双表面合成功能化方案。

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.