GOALI: Collaborative Research: Structure, Stability, and Mechanisms of Nonnative Protein Aggregate & Microparticle Formation

0931173/0932155 Roberts/Fernandez知识价值该项目旨在奠定基础，以填补对非天然微粒与可溶性聚集体形成的机械理解的关键空白，聚集体-aparticle相分离的热力学，以及稳定聚集体/颗粒并控制其形态的关键相互作用。模型蛋白质系统是α-胰凝乳蛋白酶原A（aCgn）和人免疫球蛋白γ-1（IgG 1-Fc）的Fc区。aCgn是一个充分研究的起点，基于其建立的可溶性聚集体形成机制，以及其形成聚集体和微粒的经验能力。IgG 1-Fc是一个有用的桥梁，商业上可行的蛋白质，因为它是一系列生物制药蛋白质的关键结构域，无论是基于单克隆抗体（MAbs）或融合构建体，初步数据也表明它很容易形成可溶性聚集体和颗粒。更广泛的影响该项目解决了一个长期存在的和潜在的非常高的影响在生物制药行业的问题。拟议的研究将导致更好地理解聚集和颗粒形成的机理;从而为PI实验室和其他实验室的未来努力提供基础，以合理设计和控制抗聚集性，以及控制主要类别的生物技术产品（如MAb和抗体融合蛋白）的聚集体和颗粒结构/形态。该合作研究计划涉及安进公司联合PI的重要学生指导和科学贡献，包括延长学生实习和定期团队会议。通过这些合作，包括在学术机构的工作，该项目将为PI实验室的研究生和本科生的教育和培训提供一个框架，特别关注尖端的实验和建模工具。与过去一样，弗吉尼亚大学和UD的PI将涉及来自科学和工程领域代表性不足的群体的学生。最后，一组的例子和问题将开发到本科课程，包括计算和建模活动，将本研究的各个方面。这些模块将通过圣何塞州立大学领导的基于网络的教育材料库传播给其他教师，该教育材料库由弗吉尼亚州的一名PI共同开发。

0931173/0932155Roberts/FernandezIntellectual merit This project seeks to lay a foundation to fill key gaps in mechanistic understanding of the formation of nonnative microparticles vs. soluble aggregates, the thermodynamics of aggregate-aparticle phase separation, and key interactions that stabilize aggregates / particles and control their morphology. The model protein systems are alpha-chymotrypsinogen A (aCgn) and the Fc region of human immunoglobulin gamma-1 (IgG1-Fc). aCgn is a well-studied starting point, based on its established mechanisms of soluble aggregate formation, and its empirical ability to form both aggregates and microparticles. IgG1-Fc is a useful bridge to commercially viable proteins, as it is a key domain in a range of biopharmaceutical proteins that are based on either monoclonal antibodies (MAbs) or fusion constructs, and preliminary data also indicates it readily forms both soluble aggregates and particles.Broader impactsThe project addresses a long-standing and potentially very high impact problem in the biopharmaceutical industry. The proposed research will result in an improved mechanistic understanding of aggregation and particle formation; thereby providing a basis for future efforts in the PI's laboratories and those of others to rationally design and control aggregation resistance, as well as for control of aggregate and particle structure / morphology across major classes of biotechnology products such as MAbs and antibody-fusion proteins. The collaborative research plan involves significant student mentoring and scientific contributions from the co-PI from Amgen, including extended student internships and regular team meetings. Through these collaborations, including work in the academic institutions, this project will provide a framework for the education and training of graduate and undergraduate students in the PI's laboratories, with a specific focus on cutting-edge experimental and modeling tools. As in the past, the PIs at UVA and UD will involve students drawn from underrepresented groups in science and engineering. Finally, a set of examples and problems will be developed to incorporate aspects of this research into the undergraduate curricula, including computational and modeling activities. These modules will be disseminated to other faculty via a web-based repository of educational materials led by San Jose State University that was co-developed by one PI at Virginia.