Engineering high affinity integrin binding proteins

描述（由申请人提供）：具有化学、生物化学和工程等多学科背景。这些领域将结合起来开发稳定的、高亲和力的整合素结合蛋白，用于介导肿瘤细胞粘附和血管生成。候选人将在宾夕法尼亚大学医学院的William F.退化和Joel S. Bennett的实验室做一年的导师。在此期间，申请人将在降解实验室发展肽和蛋白质设计以及有机合成方面的技能。整合素受体功能和生物检测的经验将在班尼特实验室获得。这段时间对于为该奖项的非指导部分奠定基础非常重要。剩余的奖金将用于独立的学术职位，最好是终身职位，申请人将在此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对健康和人类疾病的研究有着坚定的承诺，它的一些教师正在进行整合素和/或癌症相关的研究。申请人选择的导师在研究整合素蛋白的结构和功能方面已经建立了良好的，富有成效的合作关系。此外，在与杜邦公司合作，通过合理设计和组合库规划开发整合素特异性模拟物方面，降解教授具有丰富的专业知识。

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.