Microsystem-based Formation of Recombinant Protein Fibers

基于微系统的重组蛋白纤维的形成

• 批准号: 7894806
• 负责人: Xiaoyi Wu
• 金额: $ 18.62万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7894806
• 关键词: Affect; Aluminum; Amaze; Amino Acid Sequence; Benign; Characteristics; Controlled Study; Development; Drug Delivery Systems; Duct (organ) structure; Elastin; Engineering; Environmental Risk Factor; Fiber; Goals; Ionic Strengths; Ions; Lead; Length; Liquid substance; Lysine; Mechanics; Methodology; Microfluidic Microchips; Microfluidics; Microscope; Modeling; Molecular; Monitor; Optics; Peptide Sequence Determination; Performance; Physiological; Process; Property; Protein Biosynthesis; Protein Engineering; Proteins; Pump; Recombinant Proteins; Recombinants; Role; Scientist; Side; Silk; Site; Sodium Chloride; Solutions; Solvents; Spiders; Structure; System; Techniques; Technology; Temperature; Tissue Engineering; Water; Width; aqueous; base; cold temperature; crosslink; microsystems; mimicry; new technology; polypeptide; pressure; public health relevance; simulation; tool

DESCRIPTION (provided by applicant): The goal of the proposed project is to study protein fiber formation using an integrated microfluidic device, and to develop "green" processes for generating performance protein fibers for biomedical applications. Despite significant advances in protein synthesis and fabrication technology, spiders are still the best engineers producing extraordinarily strong silk fibers at low pressures, ambient temperatures, and with water as solvent. It has been postulated that spiders finely control the types of silk proteins, the physiological conditions of protein solutions, as well as the mechanical deformation of the silk thread in order to spin silk fibers of exceptional strength. Here, we hypothesize that the use of microsystem technology provides an important paradigm for studying the mechanisms involved in silk fiber formation and, subsequently, developing "green" processes for producing performance protein fibers. Specifically, recombinant silk-elastin-like proteins (SELPs) comprised of polypeptide sequences derived from silk and elastin will be used as a model material for the proposed project. Specific Aim 1. Determine the protein solution characteristics that influence the fiber formation and the fiber secondary structures. An integrated microfluidic system consisting of PDMS-based microchannels and an aluminum micro heater, for local temperature control, will be fabricated for studying SELP fiber formation. SELP aqueous solution will be pumped into the microfluidic system through the main channel, while salt and acidic or basic solutions will be driven through side channels to adjust the ionic strength and pH value of the SELP solution in a controlled manner. The effects of pH, ions and temperature on the fiber formation and the fiber secondary structures will be monitored using an optical microscope and characterized using a variety of spectroscopic methodologies. Specific Aim 2. Investigate the flow properties of the spinning duct that determine the fiber formation, secondary structures, and mechanical properties of SELPs. Mechanical deformation of the SELP thread will be controlled by narrowing the channel width and varying the flow rates. The resulting mechanical properties and secondary structures of SELP fibers will be characterized. In this effort, computational fluid dynamics (CFD) simulations and a constitutive analytical model will be utilized to enhance our understanding of the effects of shear/elongational flows on fiber formation, structure, and properties. Specific Aim 3. Explore the effects of the protein primary structures in modulating the formation, structure, and properties of SELP fibers. SELPs composed of varying lengths of silk-/elastin-like blocks and crosslinking sites will be synthesized, and their capability to form fibers will be examined. The influence of incorporating lysine residues, the size of silk-like blocks, as well as the ratio of silk- to elastin- like blocks on the formation, structure, and properties of SELP fibers will be investigated. PUBLIC HEALTH RELEVANCE: Microsystem-Based Formation of Recombinant Protein Fibers Spiders have amazed scientists by producing extraordinarily strong silk fibers using "green" processes at ambient temperatures, low pressures and with water as solvent. We propose to use an integrated microfluidic device as an important tool for studying the formation mechanisms of silk fibers and for developing "green" fiber fabrication techniques. Significantly, the demystification of spiders' remarkable silk spinning process and the development of "green" fabrication techniques may lead to the engineering of performance protein fibers for many biomedical applications.

描述（由申请人提供）：拟议项目的目标是研究蛋白质纤维的形成使用集成的微流体装置，并开发“绿色”的过程中产生的性能蛋白质纤维的生物医学应用。尽管在蛋白质合成和制造技术方面取得了重大进展，蜘蛛仍然是最好的工程师，可以在低压，环境温度和水作为溶剂的情况下生产非常坚固的丝纤维。据推测，蜘蛛精细地控制丝蛋白的类型、蛋白质溶液的生理条件以及丝线的机械变形，以纺出具有特殊强度的丝纤维。在这里，我们假设，微系统技术的使用提供了一个重要的范例，研究丝纤维形成的机制，随后，开发“绿色”的过程中生产的性能蛋白质纤维。具体而言，重组丝弹性蛋白样蛋白（SELP）的多肽序列来自丝绸和弹性蛋白组成将被用作模型材料的拟议项目。具体目标1。确定影响纤维形成和纤维二级结构的蛋白质溶液特性。一个集成的微流控系统，包括基于PDMS的微通道和铝微加热器，用于局部温度控制，将被制作用于研究SELP纤维的形成。SELP水溶液将通过主通道泵入微流体系统，而盐和酸性或碱性溶液将通过侧通道驱动，以受控方式调节SELP溶液的离子强度和pH值。pH值，离子和温度对纤维形成和纤维二级结构的影响将使用光学显微镜进行监测，并使用各种光谱方法进行表征。具体目标2。研究纺丝导管的流动特性，这些特性决定了SELP的纤维形成、二级结构和机械性能。SELP螺纹的机械变形将通过缩窄通道宽度和改变流速来控制。SELP纤维的机械性能和二级结构将被表征。在这项工作中，计算流体动力学（CFD）模拟和本构分析模型将被用来提高我们的理解的剪切/拉伸流动对纤维的形成，结构和性能的影响。具体目标3。探索蛋白质一级结构在调节SELP纤维的形成、结构和性质中的作用。将合成由不同长度的丝/弹性蛋白样嵌段和交联位点组成的SELP，并检查它们形成纤维的能力。将研究掺入赖氨酸残基、丝样嵌段的尺寸以及丝样嵌段与弹性蛋白样嵌段的比例对SELP纤维的形成、结构和性能的影响。 公共卫生关系：基于微系统的重组蛋白质纤维的形成蜘蛛在室温、低压和以水为溶剂的“绿色”过程中产生了非常坚固的丝纤维，这让科学家们感到惊讶。我们建议使用一个集成的微流控装置作为一个重要的工具，研究丝纤维的形成机制和发展“绿色”纤维制造技术。值得注意的是，蜘蛛卓越的吐丝过程和“绿色”制造技术的发展可能会导致许多生物医学应用的高性能蛋白质纤维的工程。

项目成果

Physical crosslinking modulates sustained drug release from recombinant silk-elastinlike protein polymer for ophthalmic applications.

• DOI: 10.1016/j.jconrel.2011.07.036
• 发表时间: 2011-12-10
• 期刊: Journal of controlled release : official journal of the Controlled Release Society
• 作者: Teng W;Cappello J;Wu X
• 通讯作者: Wu X

Modeling drug-carrier interaction in the drug release from nanocarriers.

在纳米载体中释放药物释放中的药物载体相互作用。

• DOI: 10.1155/2011/370308
• 发表时间: 2011
• 期刊: Journal of drug delivery
• 作者: Zeng L;An L;Wu X
• 通讯作者: Wu X