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.

非技术总结项目是一种集成的实验计算方法，旨在开发一类基于肽的超分子材料，作为非色谱蛋白纯化的高亲和力沉淀物。从其效率和成本效益方面，治疗蛋白与其生物资源的分离和纯化是生物制造工业制造的重要限制。尽管培养基成本较高和负载能力有限，但亲和力色谱仍然是大规模工业蛋白纯化的最广泛使用的捕获方法。由于哺乳动物细胞培养和持续过程发展，上游滴度的快速增长进一步挑战了下游制造的效率。亲和力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，合成和开发自组装的肽材料，这些肽材料可以专门绑定，选择性地捕获并有效地将蛋白质与基于生物的资源分开。此外，该项目将为各级学生参与STEM研究并体验各个学术界和行业范围的不同实验室环境，为学生提供新的教育和外展机会。技术简介旨在应对下游蛋白质制造的基于肽的亲和力降水剂开发的关键基本挑战。基本上有三个步骤在使用亲和力降水剂进行蛋白质纯化。这些是：（1）选择性捕获感兴趣的蛋白质； （2）结合诱导的相位与上清液分离； （3）蛋白质的回收。该项目包括三个特定目标，每个目标都涵盖了成功开发基于肽的超分子免疫纤维以进行非色谱蛋白纯化的关键步骤。第一个推力重点是用于选择性捕获靶标单克隆抗体（mAb）的免疫纤维的设计和合成。第二个旨在理解，确定和优化MAB结合触发的宏观相分离的条件。第三个目标集中于蛋白质恢复过程以及系统的适用性和可扩展性的评估。将开发理论和多尺度模型，以提供免疫纤维超分子设计的指导原则，以及对最大蛋白质捕获的配体呈现的策略，以捕获最大的蛋白质捕获，并帮助阐明热力学和动力学方面的体现和动力学的统计和统计的统计型构成态度的统计范围，并构成了统计的统计范围，并构成了构成构成的态度。认为值得通过基金会的智力优点和更广泛影响的评论标准来评估值得支持。