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

非技术总结本项目是一种综合的实验-计算方法，旨在开发一类基于多肽的超分子材料，作为非层析蛋白质纯化的高亲和力沉淀剂。从生物资源中分离和提纯治疗性蛋白在效率和成本效益方面是生物制品工业化生产的一个重大限制。尽管亲和层析的介质成本高，负载能力有限，但仍然是大规模工业蛋白质纯化中应用最广泛的捕获方法。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。亲和沉淀有可能克服柱子大小和配体固定所带来的层析限制。根据材料基因组计划(MGI)的目标，该项目将设计、合成和开发能够特异性结合、选择性捕获蛋白质并有效地将蛋白质从其生物资源中分离出来的自组装肽材料。此外，该项目将为所有级别的学生提供新的教育和推广机会，让他们参与STEM研究，并体验跨越学术界和行业的不同实验室环境。技术总结该项目旨在解决开发基于多肽的亲和沉淀剂用于下游蛋白质制造的关键根本挑战。亲和沉淀剂用于蛋白质纯化基本上有三个步骤。它们是：(1)选择性捕获感兴趣的蛋白质；(2)结合诱导上清液的相分离；(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基于多肽的超分子免疫纤维用于非层析蛋白质纯化的关键步骤。第一个重点是免疫纤维的设计和合成，用于选择性捕获目标单抗。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和系统适用性和可扩展性的评估上。将开发理论和多尺度模型，为免疫纤维的超分子设计和共同组装策略提供指导原则，以优化配体呈现以最大限度地捕获蛋白质，并帮助阐明免疫纤维组装和解离的热力学和动力学方面，以及结合触发的相分离机制。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.