Combinatorial Library Screening Core

组合图书馆研究核心 组合库研究核心将集中在使用几个组合库筛选 开发一系列肽或寡核苷酸适体的方法， 对肿瘤细胞、内皮细胞上的某些细胞表面受体/蛋白或内皮下细胞上的蛋白的选择性 基质或细胞内蛋白质在不同的项目中被解决。靶蛋白 包括共同犯罪中心|33用于项目1、2和4的整合素、TVA受体和HER 2受体，各种信号传导 蛋白质用于项目5、6，某些整联蛋白用于项目6。我们将使用文库筛选技术 在北卡罗来纳大学教堂山分校、杜克大学和阿贡国立大学建立的平台 实验室，包括mRNA展示，噬菌体展示和酵母双杂交技术， 选择肽适体和基于SELEX的选择核酸适体的技术。各种 将使用不同的选择方法构建和筛选肽或寡核苷酸文库。 核心项目的具体目标是：1）利用mRNA展示的10 FN 3结构域文库， 结合avps整联蛋白和TVA受体的结构域抗体模拟物; 2）使用噬菌体展示的 组合肽文库、FN 3或Top 7结构域文库和赫赛汀Fab抗体文库， 筛选HER 2结合或ccvpS结合适体或Fab片段; 3）使用以下的组合： 利用噬菌体展示进行酵母双杂交筛选，以分离结合以下的靶信号蛋白的scFv： 4）使用含有修饰的碱基（2 '-2'）的高复杂性寡核苷酸文库， 氟）来选择结合HER 2和新的肿瘤细胞标志物的RNA适体。我们将与 哈恩实验室开发用于项目5和6的基于亲和力的信号传感器。 所有选择的亲和分子将被系统地优化并表征靶结合， 特异性，并表达或合成用于应用。这些新型亲和分子的可用性 对于细胞和分子识别将在相关项目中发挥至关重要的作用。我们也将使我们的 努力促进其他癌症纳米技术中心获得这些亲和试剂 卓越

Combinatorial Library Research Core The Combinatorial Library Research Core will focus on using several combinatorial library screening approaches to develop a series of peptide or oligonucleotide aptamers that display high affinity and high selectivity for certain cell surface receptors/proteins on tumor cells, endothelial cells, or proteins on subendothelial matrix, or intracellular proteins being addressed in different projects. The target proteins include the ccv|33 integrin, TVA receptor, and HER2 receptor for projects 1, 2, and 4, various signaling proteins for projects 5, 6 and certain integrins for project 6. We will use the library screening technology platforms that have been established at UNC-Chapel Hill, Duke University, and Argonne National Laboratory, including mRNA-display, phage-display, and yeast two-hybrid based technologies for selecting peptide aptamers and SELEX based technology for selecting nucleic acid aptamers. Various peptide or oligonucleotide libraries will be constructed and screened using different selection approaches. The specific aims of the core projects are: 1) Use mRNA-displayed 10FN3 domain library to select single domain antibody mimics that bind to avps integrin and TVA receptor; 2) Use phage-displayed combinatorial peptide libraries, FN3 or Top7 domain libraries, and Herceptin Fab antibody libraries to screen for HER2-binding or ccvpS-binding binding aptamers or Fab fragments; 3) Use a combination of yeast two-hybrid screening with phage-display to isolate scFvs that bind to the target signaling proteins of interest in the cytoplasm; 4) Use high complexity oligonucleotide libraries containing modified bases (2'- fluorine) to select RNA aptamers that bind to HER2 and to novel tumor cell markers. We will work with the Hahn lab to develop affinity-based signaling sensors to be used in projects 5 and 6. All the selected affinity molecules will be systematically optimized and characterized for target binding and specificity, and expressed or synthesized for applications. The availability of these novel affinity molecules for cellular and molecular recognition will play a vital role in the related projects. We will also make our efforts to facilitate the accessibility of these affinity reagents by other Centers of Cancer Nanotechnology Excellence.