Structural Signatures for Quality Control of Proteins

蛋白质质量控​​制的结构特征

• 批准号: 6788602
• 负责人: BORIS Y ZASLAVSKY
• 金额: $ 12.06万
• 依托单位: ANALIZA, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6788602
• 关键词: 2,4 dinitrophenol; biomedical automation; biotechnology; biotherapeutic agent; drug quality /standard; high throughput technology; method development; protein folding; protein structure function; recombinant proteins; statistics /biometry

DESCRIPTION (provided by applicant): Higher-order structural information for biomolecules is an important component of quality control activities for recombinant proteins. Unfortunately, detailed structural information typically require highly complex, time-consuming experimental techniques (e.g., NMR or X-ray crystallography) that can not be considered as suitable for quality control applications. This proposal aims at developing and validating a set of techniques suitable for rapid quality control studies of biopharmaceutical products. Specifically, we aim to develop techniques that could be used to detect changes in the structure from a base state (e.g., misfolded vs. properly folded) and classify certain changes for specific protein (e.g., deamidation, altered glycosylation, mutations, etc.). Furthermore, we aim to automate the analysis methodology to increase its throughput and consistency and for ready incorporation into bioanalytical quality control services. The concept of a signature of a structure relies on combining data obtained from separate aqueous two-phase partitioning experiments that are sensitive to small structural modifications, with mathematical analysis techniques that make the signature highly specific. Signatures contain much less information than the underlying structure itself and are thus constructed specifically for each application. Phase I research work is centered on providing feasibility data to demonstrate the generality of signatures with for one or more biopharmaceutical products with respect to different structure modifications typical of and important for quality control. Phase I would also provide initial data concerning the proper design of experimental techniques and preferred mathematical and statistical methods for constructing signatures on a routine basis. Phase II work would expand upon this work to further generalize and validate the methodology for multiple proteins with multiple potential degradation pathways, and on incorporating the techniques into an automated signature workstation for rapid low-cost determining and deciphering of structural information of production samples. This proposal further extends previous work related to quality control of bioterrorism-related vaccine products to provide the depth and breadth of analysis and data necessary for adaptation by industry.

描述(申请人提供)：生物分子的高阶结构信息是重组蛋白质质量控制活动的重要组成部分。不幸的是，详细的结构信息通常需要高度复杂、耗时的实验技术(例如，核磁共振或X射线结晶学)，而不能被认为适合于质量控制应用。该建议旨在开发和验证一套适用于生物制药产品快速质量控制研究的技术。具体地说，我们的目标是开发一种技术，可以用来检测碱基状态下的结构变化(例如，错误折叠与正确折叠)，并对特定蛋白质的某些变化进行分类(例如，脱酰胺、糖基化改变、突变等)。此外，我们的目标是实现分析方法的自动化，以增加其吞吐量和一致性，并随时纳入生物分析质量控制服务。结构签名的概念依赖于将从对微小结构修改敏感的单独的双水相分配实验中获得的数据与使签名高度专一的数学分析技术相结合。签名包含的信息比底层结构本身少得多，因此是专门为每个应用程序构造的。第一阶段的研究工作集中于提供可行性数据，以证明一种或多种生物制药产品在不同结构修改方面签名的普遍性，这是典型的和对质量控制重要的。第一阶段还将提供关于实验技术的适当设计的初始数据，以及用于常规构造签名的优选的数学和统计方法。第二阶段的工作将在这项工作的基础上进一步推广和验证具有多种潜在降解途径的多种蛋白质的方法学，并将这些技术纳入一个自动签名工作站，以便以低成本快速确定和破译生产样品的结构信息。这项建议进一步扩展了以前与生物恐怖主义相关疫苗产品质量控制有关的工作，以提供行业适应所需的分析和数据的深度和广度。