Protein purification by selective coacervation

通过选择性凝聚纯化蛋白质

• 批准号: 0966923
• 负责人: Paul Dubin
• 金额: $ 30万
• 依托单位: University of Massachusetts Amherst
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0966923&HistoricalAwards=false
• 关键词: Protein; purification; selective; coacervation

Protein drugs are important in the treatment of diabetes, leukemia, hemophilia, Alzheimer's and Gaucher disease, and osteoporosis. They are also important therapeutic agents in organ replacement, tissue regeneration and wound repair, regardless of whether resulting from disease, aging or battle related injury. The technologies for large scale protein separation needed to produce these drugs have lagged behind molecular biology, and now are responsible for most of their cost. The use of generally recognized as safe (GRAS) polymers (polyelectrolytes) are investigated to isolate and concentrate the target proteins from the cell crush or lysate in which they are formed. The process by which this happens is a spontaneous separation into two liquids: the more dense phase contains the polymer and the target protein in high concentration. The goal of this work is to understand the principles by which this complex coacervation can be optimized by (a) the selection of the polymer, (b) the choice of the conditions for effecting the coacervation, and (c) the method of removing polymer from the target protein.The interdisciplinary coupling of bioengineering, bioanlytical, and polymer physical chemistry proposed provide an example of the unification of perspectives appropriate to progress in this field. The PIs take a pro-active role in involving members of underrepresented groups in research. Both gender and racial minorities make up a significant proportion of their groups. There are currently nine women in the two groups (1 postdoc, 4 graduates, 3 undergraduates, 1 high school senior), three of whom are involved in work related to this proposal (protein polyelectrolyte coacervation). There is active participation in an NSF REU and IGERT grants for promoting undergraduate research. Students working on this project will obtain training in areas of great importance to biotechnology and pharmaceutical innovation and applications.

蛋白质药物在治疗糖尿病、白血病、血友病、阿尔茨海默病和戈谢病以及骨质疏松症中是重要的。它们也是器官置换、组织再生和伤口修复中的重要治疗剂，无论是否由疾病、衰老或与战斗有关的损伤引起。生产这些药物所需的大规模蛋白质分离技术落后于分子生物学，现在承担了大部分成本。 研究了使用公认安全（GRAS）聚合物（聚电解质）从形成靶蛋白的细胞破碎物或裂解物中分离和浓缩靶蛋白。发生这种情况的过程是自发分离成两种液体：更致密的相含有高浓度的聚合物和靶蛋白。本工作的目标是理解通过（a）聚合物的选择，（B）实现凝聚的条件的选择，和（c）从靶蛋白去除聚合物的方法来优化这种复合凝聚的原理。和高分子物理化学的提出提供了一个例子，统一的观点，适当的进展，在这一领域。 PI在让代表性不足的群体成员参与研究方面发挥积极作用。性别和种族少数群体在其群体中占很大比例。目前，这两个小组共有9名妇女（1名博士后、4名研究生、3名本科生、1名高中生），其中3人参与了与该提案有关的工作（蛋白质凝聚）。积极参与NSF REU和IGERT赠款，以促进本科生研究。从事该项目的学生将获得对生物技术和制药创新和应用非常重要的领域的培训。