Ultrahigh Performance Non-Denaturing Protein Chromatography Columns

超高性能非变性蛋白质色谱柱

• 批准号: 9202251
• 负责人: MARY J. WIRTH
• 金额: $ 13.97万
• 依托单位: BIOVIDRIA, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-16 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9202251
• 关键词: Address; Adsorption; Adverse effects; Anions; Back; Cations; Characteristics; Charge; Chromatography; Column Chromatography; Development; Drug usage; Effectiveness; Future; Goals; Growth; Heterogeneity; Hydrophobic Interactions; Length; Liquid Chromatography; Longitudinal Studies; Malignant Neoplasms; Marketing; Measures; Modification; Monoclonal Antibodies; Outcome; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Polymers; Procedures; Process; Proteins; Reaction Time; Reproducibility; Research; Resolution; Silanes; Silicon Dioxide; Stainless Steel; Surface; Time; Water; Work; efficacy testing; flexibility; immunogenicity; improved; innovation; manufacturing process; molecular scale; monolayer; particle; polymerization; pressure; scale up; statistics; time use

PROJECT SUMMARY The proposed product is an analytical scale, UHPLC column for separating protein drugs in native form, with the Phase I proposal focused on anion exchange chromatography as the separation mechanism. The bonded phase is made of polymer chains, in this case bearing weak anion exchange groups. The study will be expanded to cation exchange and hydrophobic interaction chromatography in future Phase II work. These non-denaturing separations are essential in the development of protein drugs because they allow the separated components to be tested for efficacy, immunogenicity and mechanism in phamacological research. Non- denaturing chromatography columns for protein drugs use polymeric bonded phases because these impart the molecular-scale flexibility needed for fast adsorption and desorption. The overall market size for non-denaturing chromatography columns is on the order of $1B, and the market leaders include Thermo-Dionex. Agilent, Waters and Sepax. The problems with current columns are two-fold: the separations are relatively low in resolution and the columns have short lifetimes. Preliminary results show both resolution column lifetime can be improved by an innovative manufacturing process. The latter considers that resolution and lifetime are limited because polymers are compressible, and the cycling between high pressure and ambient pressure through routine use of the column progressively disturbs the packing to degrade the separation over time. The proposed columns will be made by first tightly packing incompressible silica particles to give a pressure-invariant packing. Then the polymer bonded phase will be grown inside the column from initiators on the surfaces of the silica particles, forming a uniform bonded phase. The proposed work entails two specific aims. The first aim is to develop an automated procedure for in-column growth of the polymeric bonded phase. The second aim is to compare the column performance with those of leading commercial columns with respect to resolution of monoclonal antibodies having known charge heterogeneity and column lifetime. The outcome of the work will establish the feasibility of commercializing a line of non-denaturing chromatography columns for protein drugs. The concept will be extended in the future Phase II research to address scale-up issues in manufacturing and to develop other non-denaturing separations, including cation chromatography and hydrophobic interaction chromatography.

项目摘要 拟定产品是一种分析规模的UHPLC色谱柱，用于分离蛋白质药物， 天然形式，第一阶段的建议集中在阴离子交换色谱作为 分离机制粘合相由聚合物链组成，在这种情况下， 弱阴离子交换基团。该研究将扩展到阳离子交换和疏水 相互作用色谱法在未来的第二阶段工作。这些非变性分离是 在蛋白质药物的开发中至关重要，因为它们允许分离的组分 在药理学研究中进行有效性、免疫原性和机制测试。非 用于蛋白质药物的变性层析柱使用聚合物键合相， 这些赋予了快速吸附和解吸所需的分子尺度的灵活性。的 非变性色谱柱的总体市场规模约为10亿美元， 市场领导者包括Thermo-Dionex。Agilent、沃茨和Sepax。当前的问题 色谱柱是双重的：分离的分辨率相对较低，色谱柱具有 短寿命初步结果表明，两个分辨率列寿命可以提高一个 创新的制造工艺。后者认为分辨率和寿命有限 因为聚合物是可压缩的，并且高压和环境压力之间的循环 通过柱的常规使用的压力逐渐地扰动填料以降解 随着时间的推移分离。建议的列将由第一紧密包装不可压缩 二氧化硅颗粒以提供压力不变的填料。然后，聚合物粘结相将是 在柱内从二氧化硅颗粒表面上的引发剂生长，形成均匀的 键合相拟议的工作有两个具体目标。第一个目标是发展一个 用于聚合物键合相的柱内生长的自动化程序。第二个目标是 将色谱柱性能与领先的商业色谱柱性能进行比较， 具有已知电荷异质性和柱寿命的单克隆抗体的分离度。 这项工作的结果将建立商业化的非变性线的可行性， 用于蛋白质药物的色谱柱。这一概念将在未来的第二阶段得到扩展 研究以解决制造业的规模扩大问题，并开发其他非变性 分离，包括阳离子色谱和疏水相互作用色谱。