Improved Protein Separations with Superficially Porous Particles

使用表面多孔颗粒改进蛋白质分离

• 批准号: 8978832
• 负责人: Barry E Boyes
• 金额: $ 21.93万
• 依托单位: ADVANCED MATERIALS TECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8978832
• 关键词: Address; Area; Biological; Biological Process; Biological Products; Biological Sciences; Biomedical Research; Caliber; Characteristics; Chemistry; Complex; Development; Dimensions; Disease; Evaluation; Generations; Genetic; Glycopeptides; Glycoproteins; Goals; High Pressure Liquid Chromatography; Hour; Individual; Knowledge; Laboratories; Lead; Liquid Chromatography; Liquid substance; Literature; Marketing; Mass Spectrum Analysis; Measurement; Methods; Modification; Outcome; Particle Size; Peptides; Performance; Pharmacologic Substance; Polysaccharides; Post-Translational Protein Processing; Property; Proteins; Quality Control; Relative (related person); Research; Resolution; Safety; Sampling; Science; Silicon Dioxide; Structure; Surface; System; Technology; Therapeutic Agents; Therapeutic Intervention; Thick; Time; Work; analytical method; design; improved; interest; meetings; new technology; novel; particle; product development; protein expression; public health relevance

 DESCRIPTION (provided by applicant): Life sciences research and other critical bioanalytical applications would strongly benefit from faster and higher resolution liquid chromatographic (HPLC) separations of larger molecules including proteins, peptides, glycopeptides, and glycans. Many HPLC separations of biomedical interest, such as resolution of protein mixtures, require an hour or more, and may not completely resolve all of the components. Due to the complexity of biological samples, there are many examples in the current research literature that combine multiple dimensions of HPLC separations, often with online mass spectroscopy (MS). Although very time consuming, this approach has yielded useful information on protein expression and post-translational modifications, the biological processes underlying protein expression and modifications, and the effects thereon of development, disease state, and various environmental and genetic factors. In other cases, such as the development of protein therapeutic agents, many closely related protein molecules may be present, but precise knowledge of the individual identities and quantities of each could be required for assurance of safety and efficacy of the therapeutic agent. In either circumstance, high efficiency LC separations are a fundamental part of the analytical systems, and the time to achieve high resolution separations of complex biological samples is a great bottleneck. This proposal describes an approach to improve the separation efficiency of HPLC by extending the use of superficially porous particle (SPP) technologies to smaller size particles, with optimized characteristics, providing significantly faster and higher resolution separations. Our objectives are to create 2.0 µm diameter and smaller SPP silica particles, optimize their materials properties, and to load these materials efficiently into HPLC column formats. The goals of the proposed work would yield very high performance chromatographic products, greater than those currently available, to be applied broadly in analysis of complex biological samples, pharmaceutical and biopharmaceutical applications, in fact in any current application that uses HPLC methods for larger biological molecule analyses. The separations technology described will directly lead to useful products for which there is a significant technical and market demand.

 描述（由申请人提供）：生命科学研究和其他关键生物分析应用将极大受益于对较大分子（包括蛋白质、肽、糖肽和聚糖）进行更快、更高分辨率的液相色谱（HPLC）分离。许多具有生物医学意义的HPLC分离，如蛋白质混合物的分离，需要一个小时或更长时间，并且可能无法完全分离所有组分。由于生物样品的复杂性，在当前的研究文献中有许多实例将联合收割机的多个维度的HPLC分离（通常与在线质谱（MS））相结合。虽然非常耗时，但这种方法已经产生了关于蛋白质表达和翻译后修饰、蛋白质表达和修饰的生物学过程以及发育、疾病状态和各种环境和遗传因素对蛋白质表达和修饰的影响的有用信息。在其他情况下，例如蛋白质治疗剂的开发，可能存在许多密切相关的蛋白质分子，但可能需要精确了解每种蛋白质分子的个体身份和数量，以确保治疗剂的安全性和有效性。在任一情况下，高效LC分离是分析系统的基本部分，并且实现复杂生物样品的高分辨率分离的时间是一个很大的瓶颈。该提案描述了一种通过将表面多孔颗粒（SPP）技术的使用扩展到具有优化特性的较小尺寸颗粒来提高HPLC分离效率的方法，从而提供显著更快和更高分辨率的分离。我们的目标是制造直径为2.0 µm和更小的SPP二氧化硅颗粒，优化其材料特性，并将这些材料有效地加载到HPLC柱格式中。所提出的工作的目标将产生非常高性能的色谱产品，大于目前可用的，被广泛应用于复杂的生物样品，制药和生物制药应用的分析，事实上，在任何目前的应用，使用HPLC方法进行较大的生物分子分析。所描述的分离技术将直接导致有用的产品，对于这些产品存在显著的技术和市场需求。