Fabrication of Novel Biomimetic Polymers Using Combinatorial Peptide Screening

利用组合肽筛选制备新型仿生聚合物

• 批准号: 7227548
• 负责人: CHRISTINE E SCHMIDT
• 金额: $ 24.91万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-07 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7227548
• 关键词: Adhesives; Affinity; Anions; Atomic Force Microscopy; Bacteriophages; Binding; Biocompatible Materials; Biological; Biological Assay; Biomimetics; Biosensor; Calorimetry; Capsid Proteins; Cells; Characteristics; Charge; Chemicals; Chlorine; Complex; Computer Simulation; Count; Coupling; Devices; Diagnostic; Drug Delivery Systems; Engineering; Enzymes; Fluorescamine; Goals; Growth Factor; Histology; Image; Immune response; Immunofluorescence Immunologic; Implant; In Vitro; Inflammation; Ions; Libraries; Life; Lysine; Measurement; Measures; Methods; Modification; Molecular Weight; Monitor; Natural regeneration; Nature; Nerve Growth Factor 1; Nerve Growth Factor Pathway; Nerve Growth Factors; Nerve Regeneration; Peptide Phage Display Library; Peptides; Phage Display; Pharmaceutical Preparations; Polymers; Property; Proteins; RGD (sequence); Range; Rattus; Reaction; Regenerative Medicine; Relative (related person); Research Personnel; Screening procedure; Serum; Site; Surface; Techniques; Testing; Thermodynamics; Tissue Engineering; Tissues; Titrations; United States Food and Drug Administration; Variant; analog; biomaterial compatibility; chemical group; combinatorial; design; implantation; in vivo; interest; models and simulation; molecular dynamics; monomer; novel; physical property; polymerization; polypyrrole; programs; protein aminoacid sequence; response; retinal rods; size; subcutaneous

DESCRIPTION (provided by applicant): The goal of these studies is to develop and characterize unique synthetic polymer-biological molecule composites for biomedical applications. As part of preliminary studies, combinatorial peptide phage display libraries and biopanning techniques have been used to select a unique peptide sequence ("T59") that specifically and tightly binds directly to chlorine-doped polypyrrole (PPyCl), an electrically conductive polymer that has shown promise in biomedical applications, such as nerve regeneration. In the proposed studies, the binding affinity and stability of the T59 peptide interaction with PPyCl (both in vitro and in vivo) and the nature of this interaction will be investigated. This information will contribute to the understanding of surface interactions and biomaterials modification strategies, and is critical for effectively applying these sequences for either in vitro (e.g., biosensor) or in vivo (e.g., tissue engineering) applications. PPyCl will also be functionalized with large biomolecules (e.g., NGF) to illustrate the versatility and utility of this approach. These goals will be accomplished in the following Specific Aims: (1) Study the in vivo response to PPyCl modified with T59; (2) quantitatively analyze, using fluorescamine protein assays, atomic force microscopy, and isothermal titration calorimetry, the binding of T59 to PPyCl; (3) use chemical and polymer analogs in conjunction with peptide variants (designed using modeling and simulations of binding energetics) to study the mechanism of interaction between T59 and PPyCl; and (4) study the ability to attach large biomolecules (i.e., nerve growth factor or NGF) to PPyCl via the T59 peptide. Overall, these studies will explore an alternate approach for modifying synthetic polymers for tissue engineering applications. By selecting and identifying unique peptide sequences that interact with high affinity to synthetic polymers, one can easily modify the polymer surfaces using those peptides (i.e., peptides can be synthesized with the polymer binding sequence on one end, and a sequence that binds to cells, drugs, growth factors, etc. on the other end). This strategy for surface modification could serve as a versatile method to develop bioactive materials using existing polymers (including those that are already FDA approved and/or those polymers that lack functional chemical groups for coupling reactions, like PPyCl), without changing the bulk properties of the materials.

描述（由申请人提供）：这些研究的目标是开发和表征用于生物医学应用的独特的合成聚合物-生物分子复合材料。作为初步研究的一部分，组合肽噬菌体展示文库和生物淘选技术已用于选择独特的肽序列（“T59”），该序列特异性且紧密地直接与氯掺杂聚吡咯（PPyCl）结合，这是一种导电聚合物，在生物医学应用（例如神经再生）中显示出前景。 在拟议的研究中，将研究 T59 肽与 PPyCl 相互作用的结合亲和力和稳定性（体外和体内）以及这种相互作用的性质。这些信息将有助于理解表面相互作用和生物材料修饰策略，并且对于有效地将这些序列应用于体外（例如生物传感器）或体内（例如组织工程）应用至关重要。 PPyCl 还将用大生物分子（例如 NGF）进行功能化，以说明该方法的多功能性和实用性。这些目标将通过以下具体目标来实现：（1）研究T59修饰的PPyCl的体内反应； (2)利用荧光胺蛋白测定、原子力显微镜和等温滴定量热法定量分析T59与PPyCl的结合； (3) 使用化学和聚合物类似物与肽变体（使用结合能量学建模和模拟设计）来研究T59和PPyCl之间的相互作用机制； (4) 研究通过 T59 肽将大生物分子（即神经生长因子或 NGF）附着到 PPyCl 上的能力。 总体而言，这些研究将探索一种修改组织工程应用的合成聚合物的替代方法。通过选择和识别与合成聚合物具有高亲和力相互作用的独特肽序列，人们可以使用这些肽轻松修饰聚合物表面（即，可以合成肽，其一端具有聚合物结合序列，另一端具有与细胞、药物、生长因子等结合的序列）。这种表面改性策略可以作为一种通用方法，使用现有的聚合物（包括那些已经获得 FDA 批准的聚合物和/或那些缺乏用于偶联反应的功能性化学基团的聚合物，如 PPyCl）来开发生物活性材料，而不改变材料的整体特性。