Combinatorial Library Screening Core

组合图书馆研究核心 组合文库研究核心将重点利用几个组合文库筛选 开发一系列具有高亲和力和高亲和力的多肽或寡核苷酸适配子的方法 肿瘤细胞、内皮细胞或内皮下细胞上某些细胞表面受体/蛋白的选择性 基质，或在不同项目中处理的细胞内蛋白质。靶蛋白 包括CCV|33整合素、TVA受体和HER2受体项目1、2和4，各种信号转导 项目5、6的蛋白质和项目6的某些整合素。我们将使用文库筛选技术 已经在北卡罗来纳大学-教堂山、杜克大学和阿贡国家大学建立的平台 实验室，包括基于mRNA展示、噬菌体展示和酵母双杂交的技术 选择肽适配子和基于SELEX的核酸适配子选择技术。五花八门 将使用不同的选择方法构建和筛选多肽或寡核苷酸文库。 核心项目的具体目标是：1)使用mRNA展示的10FN3结构域文库来选择单个 与AVP整合素和TVA受体结合的结构域抗体；2)使用噬菌体展示 组合多肽文库、FN3或Top7结构域文库和Herceptin Fab抗体库 筛选HER2结合或ccvpS结合的适体或Fab片段；3)使用 噬菌体展示酵母双杂交技术筛选与猪瘟病毒靶信号蛋白结合的单链抗体 对细胞质感兴趣；4)使用含有修饰碱基(2‘-)的高复杂性寡核苷酸文库 氟)选择与HER2结合的RNA适配子和新的肿瘤细胞标记物。我们将与 哈恩实验室开发基于亲和力的信号传感器，用于项目5和6。 所有选定的亲和分子都将进行系统的优化和表征，以进行靶向结合和 特异性，并为应用而表达或合成。这些新型亲和分子的可用性 因为细胞和分子识别将在相关项目中发挥至关重要的作用。我们还将使我们的 促进其他癌症纳米技术中心获取这些亲和试剂的努力 太棒了。

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.