Elastin Fusion Proteins

弹性蛋白融合蛋白

• 批准号: 7856274
• 负责人: Ashutosh Chilkoti
• 金额: $ 13.5万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-17 至 2010-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7856274
• 关键词: Affinity; Affinity Chromatography; Area; Behavior; Binding; Biocompatible Materials; Biological; Biomedical Engineering; Biomimetics; Biotechnology; C-terminal; Chemicals; Chimeric Proteins; Complex; Complex Mixtures; Development; Elastin; Ensure; Exhibits; Funding; Genes; Goals; Heating; Immobilization; Immunoglobulin G; Investigation; Methods; Modification; Molecular; Molecular Models; Molecular Weight; Phase Transition; Polymers; Protein Binding Domain; Protein Microchips; Proteins; Recombinant Proteins; Recombinants; Recovery; Relative (related person); Research; Research Personnel; Resolution; Set protein; Side; Site-Directed Mutagenesis; Sodium Chloride; Solubility; Solutions; Solvents; Stimulus; Structure; Surface; Surface Properties; System; Technology; Temperature; Testing; Therapeutic; analog; aqueous; base; chemical property; feeding; immunoaffinity chromatography; interest; micro-total analysis system; molecular modeling; nanocarrier; overexpression; polypeptide; programs; prolylglycine; prolylvaline; protein purification; receptor; response; therapeutic protein

DESCRIPTION (provided by applicant): The overall objective of this competing R01 renewal is the continued development of the Elastin Fusion Protein technology as a genetically encodable, biomimetic analog of -"smart" protein-polymer conjugates. The overall objective is motivated by the rationale that simple and convenient methods to modulate the physico-chemical properties of proteins -most notably their solubility in solution and binding to a surface- in response to an external trigger have immediate application in many areas of Bioengineering and Biotechnology such as protein purification, affinity capture, protein chips, lab-on-a-chip bioanalysis and in the development of environmentally responsive biomaterials and nanocarriers for delivery of protein therapeutics. The objective of the proposed research will be accomplished by the gene-level fusion of different recombinant proteins to a stimulus-responsive elastin-like polypeptide (ELP) tag to create ELP fusion proteins. The overall hypothesis of this research -validated for a set of proteins in the previous funding period- is that environmentally sensitive and reversible solubility and binding to a surface can be imparted to any soluble protein of interest by N-or C-terminal fusion to an ELP sequence. The proposed research has two research foci: the first, "fundamental focus", seeks to discover the biophysical basis of the effect that fusion of proteins has on the inverse transition behavior of an ELP. The molecular level understanding of the inverse transition behavior of ELP fusion proteins will directly feed into the second, "technological focus" that will develop an ensemble of applications that exploit the inverse transition behavior of ELP fusion proteins. The second focus consists of three related technological applications: (1) optimization of the expression and purification of ELP fusion proteins in a bacterial expression system; (2) affinity capture of biomolecules by an ELP fusion protein followed by their separation by inverse transition cycling; and (3) "direct, on chip", reversible capture of ELP fusion proteins from solution on surfaces functionalized with ELPs.

描述（由申请人提供）：这种竞争R01更新的总体目标是弹性蛋白融合蛋白技术的持续开发是一种基因编码，仿生的类似物 - “智能”蛋白质 - 聚合物偶联物。 The overall objective is motivated by the rationale that simple and convenient methods to modulate the physico-chemical properties of proteins -most notably their solubility in solution and binding to a surface- in response to an external trigger have immediate application in many areas of Bioengineering and Biotechnology such as protein purification, affinity capture, protein chips, lab-on-a-chip bioanalysis and in the development of environmentally responsive用于蛋白质疗法的生物材料和纳米载体。拟议研究的目的将通过不同重组蛋白与刺激反应性弹性蛋白样多肽（ELP）标签的基因级融合来实现，以创建ELP融合蛋白。这项研究对一组蛋白质在先前的资金期间进行了验证的总体假设是，环境敏感和可逆的溶解度以及与表面的结合可以通过N-OR C末端融合到ELP序列的任何感兴趣的蛋白质。拟议的研究有两个研究重点：第一个“基本重点”试图发现蛋白质融合对ELP的反向过渡行为的影响的生物物理基础。对ELP融合蛋白的反向过渡行为的分子水平理解将直接进入第二个“技术焦点”，该“技术重点”将开发出利用ELP融合蛋白的反向过渡行为的应用集合。第二个焦点由三种相关的技术应用组成：（1）在细菌表达系统中优化ELP融合蛋白的表达和纯化； （2）通过ELP融合蛋白对生物分子的亲和力捕获，然后通过反过渡循环分离； （3）“直接，在芯片上”，可逆地捕获ELP融合蛋白从溶液中用ELP官能化的表面上的溶液。