Collaborative Research: DMREF: GOALI: High-Affinity Supramolecular Peptide Materials for Selective Capture and Recovery of Proteins

合作研究：DMREF：GOALI：用于选择性捕获和回收蛋白质的高亲和力超分子肽材料

• 批准号: 2119686
• 负责人: Monica Olvera
• 金额: $ 60万
• 依托单位: Northwestern University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2119686&HistoricalAwards=false
• 关键词: Collaborative; Research; DMREF; GOALI; Affinity

NON-TECHNICAL SUMMARYThis project is an integrated experimental-computational approach that aims to develop a class of peptide-based supramolecular materials as high-affinity precipitants for non-chromatographic protein purification. The separation and purification of therapeutic proteins from their biological resources are a significant limitation for industrial manufacturing of biologics in terms of their efficiency and cost-effectiveness. Despite the high media cost and limited loading capacity, affinity chromatography remains the most widely used capture method for large-scale industrial protein purification. The rapid growth of upstream titers, due to advancements in mammalian cell culture and continuous process development, has further challenged the efficiency of downstream manufacturing. Affinity precipitation can potentially overcome the chromatography limitations associated with column size and ligand immobilization. In line with the goals of the Materials Genome Initiative (MGI), this project will design, synthesize, and develop self-assembling peptide materials that can specifically bind, selectively capture, and effectively separate proteins from their bio-based resources. In addition, this project will foster new educational and outreach opportunities for students at all levels to participate in STEM research and to experience different laboratory settings that range across academia and industry.TECHNICAL SUMMARYThis project aims to address the key fundamental challenges in the development of peptide-based affinity precipitants for downstream protein manufacturing. There are essentially three steps in the use of affinity precipitants for protein purification. These are the: (1) selective capture of proteins of interest; (2) binding-induced phase separation from supernatant; and (3) recovery of proteins. The project includes three specific aims, each covering a key step toward the successful development of peptide-based supramolecular immunofibers for non-chromatographic protein purification. The first thrust focuses on the design and synthesis of immunofibers for selective capture of target monoclonal antibodies (mAbs). The second is intended to understand, determine, and optimize the conditions for mAb binding-triggered macroscopic phase separation. The third aim centers on the protein recovery processes and the assessment of system applicability and scalability. Theory and multiscale models will be developed to provide guiding principles for the supramolecular design of immunofibers and the co-assembly strategies to optimize the ligand presentation for maximal protein capture, and to help elucidate the thermodynamic and kinetic aspects of immunofiber assembly and dissociation, as well as the binding-triggered phase separation mechanisms.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

非技术摘要该项目是一种综合实验计算方法，旨在开发一类基于肽的超分子材料作为非色谱蛋白质纯化的高亲和力沉淀剂。从生物资源中分离和纯化治疗性蛋白质是生物制剂工业制造的效率和成本效益的重大限制。尽管介质成本高且上样量有限，亲和层析仍然是大规模工业蛋白质纯化中最广泛使用的捕获方法。由于哺乳动物细胞培养的进步和持续的工艺开发，上游滴度的快速增长进一步挑战了下游制造的效率。亲和沉淀可以潜在地克服与柱尺寸和配体固定相关的色谱限制。根据材料基因组计划（MGI）的目标，该项目将设计、合成和开发自组装肽材料，这些材料可以特异性结合、选择性捕获并有效地将蛋白质与其生物基资源分离。此外，该项目将为各级学生提供新的教育和推广机会，让他们参与 STEM 研究并体验学术界和工业界的不同实验室环境。技术摘要该项目旨在解决肽开发中的关键基本挑战用于下游蛋白质制造的基于亲和力的沉淀剂。使用亲和沉淀剂进行蛋白质纯化基本上分为三个步骤。这些是：（1）选择性捕获感兴趣的蛋白质； (2) 结合诱导的上清液相分离； (3)蛋白质的回收。该项目包括三个具体目标，每个目标都涵盖了成功开发用于非色谱蛋白质纯化的基于肽的超分子免疫纤维的关键一步。第一个重点是用于选择性捕获目标单克隆抗体（mAb）的免疫纤维的设计和合成。第二个目的是了解、确定和优化 mAb 结合触发的宏观相分离的条件。第三个目标集中在蛋白质回收过程以及系统适用性和可扩展性的评估。将开发理论和多尺度模型，为免疫纤维的超分子设计和共组装策略提供指导原则，以优化配体呈现以实现最大程度的蛋白质捕获，并帮助阐明免疫纤维组装和解离的热力学和动力学方面作为结合触发的相分离机制。该奖项反映了 NSF 的法定使命，并且通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。