Engineering high affinity integrin binding proteins

工程化高亲和力整合素结合蛋白

• 批准号: 6712379
• 负责人: JENNIFER R COCHRAN
• 金额: $ 13.08万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-10 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6712379
• 关键词: angiogenesis; binding proteins; biotechnology; carcinogenesis inhibitor; cell adhesion; chemical registry /resource; chemical stability; crosslink; cyclic peptides; drug design /synthesis /production; flow cytometry; high performance liquid chromatography; immunologic substance resource /registry /referral center; integrins; mass spectrometry; matrix assisted laser desorption ionization; metastasis; protein binding; protein engineering; protein folding; protein structure function; receptor coupling; tissue support frame; vascular endothelium; yeast two hybrid system

DESCRIPTION (provided by applicant): The candidate has a multidisciplinary background in chemistry, biochemistry and engineering. These fields will be combined to develop stable, high affinity integrin binding proteins for mediating cell adhesion and angiogenesis in cancer. The candidate will spend a year in a mentored position in the laboratories of William F. DeGrade and Joel S. Bennett at the University of Pennsylvania School of Medicine. During this period, the applicant will develop skills in peptide and protein design, and organic synthesis while in the DeGrade lab. Experience in integrin receptor function and biological assays will be gained in the Bennett lab. This period will be important in establishing the groundwork for the unmentored portion of the award. The remainder of the award will be spent in an independent, preferrably tenure-track, academic position, where the applicant will build upon this additional expertise to develop biomolecules for cancer therapeutics and tumor imaging applications. The University of Pennsylvania School of Medicine has a strong commitment to research in health and human disease, and several of its faculty are conducting integrin and/or cancer related research. The mentors chosen by the applicant have well-established, productive collaborations in studying the structure and function of integrin proteins. In addition, Prof. DeGrade has much expertise in development of integrin-specific mimics through rational design and combinatorial library panning in collaboration with DuPont. Integrin-specific peptide mimics and antibodies have shown much therapeutic promise in the inhibition of angiogenesis and metastases in tumors, but can benefit greatly from stability and affinity maturation. The mentored period of the award will involve incorporation of integrin-specific peptide motifs into constrained molecular scaffolds to increase stability and binding affinity. These scaffolds will provide the framework for a portion of the unmentored period of the award, where integrin binding proteins will be engineered to even higher affinity using directed molecular evolution by yeast surface display. In addition, the mentored period of the award will focus on the design and synthesis of a series of novel peptide-based crosslinking reagents. These compounds will be used for creating multivalent integrin-specific peptides and proteins for high avidity binding and enhanced antagonism. In the unmentored period of the application, a nonimmune human library of scFv antibody fragments displayed on the surface of yeast will be used to isolate a panel of integrin-specific binders. These antibodies will be characterized for their ability to inhibit RGD-mediated binding and cell adhesion. Promising antibodies will be engineered to ultra high affinity and stability using yeast surface display to develop potent integrin antagonists.

应聘者描述(由申请人提供)：应聘者具有化学、生物化学和工程等多学科背景。这些领域将结合在一起，开发稳定的、高亲和力的整合素结合蛋白，用于介导癌症中的细胞黏附和血管生成。候选人将在宾夕法尼亚大学医学院威廉·F·德格莱德和乔尔·S·班尼特的实验室担任一年的导师职位。在此期间，申请者将在降解实验室中发展多肽和蛋白质设计以及有机合成方面的技能。贝内特实验室将获得整合素受体功能和生物检测方面的经验。这段时间对于为奖项的无指导部分奠定基础将是重要的。该奖项的其余部分将在一个独立的、最好是终身教职的学术职位上度过，申请者将在这一额外专业知识的基础上开发用于癌症治疗和肿瘤成像应用的生物分子。宾夕法尼亚大学医学院致力于健康和人类疾病的研究，其几名教员正在进行整合素和/或癌症相关研究。申请者选择的导师在研究整合素蛋白质的结构和功能方面有良好的、富有成效的合作。此外，德格德教授在通过合理设计和与杜邦合作的组合文库淘选开发整合素特异性模拟药物方面拥有丰富的专业知识。 整合素特异性多肽模拟物和抗体在抑制肿瘤血管生成和转移方面显示出很大的治疗前景，但可以从稳定性和亲和力成熟中受益匪浅。该奖项的指导期将包括将整合素特定的多肽基序整合到受限制的分子支架中，以增加稳定性和结合亲和力。这些支架将为该奖项的非指导期的一部分提供框架，其中整合素结合蛋白将通过酵母表面展示的定向分子进化被设计成更高的亲和力。此外，该奖项的导师期将专注于一系列新型多肽基交联剂的设计和合成。这些化合物将用于产生多价整合素特异性多肽和蛋白质，以实现高亲和力结合和增强拮抗作用。在应用的非指导期，展示在酵母表面的单链抗体片段的非免疫性人类文库将被用来分离一组整合素特异性结合蛋白。这些抗体的特征是它们能够抑制RGD介导的结合和细胞黏附。有希望的抗体将被设计成超高亲和力和稳定性，使用酵母表面展示来开发有效的整合素拮抗剂。