Polypeptide Multilayer Nanofilms for Synthetic Biomatrix Development

描述（由申请人提供）：SBIR第一阶段项目将确定将多肽多层纳米膜技术应用于合成生物基质开发和商业化的可行性。这些纳米膜是一种用于细胞和组织培养的表面涂层。合成生物基质材料的直接体外应用包括基础医学研究、诊断测试、高通量药物筛选、细胞表型选择、生物制剂生产和各种治疗性细胞技术的开发的许多领域。全球合成生物基质市场的年价值估计接近10亿美元。组织中任何不是细胞的部分都被称为细胞外基质（ECM）。在活体中，ECM包围细胞并影响细胞形状、增殖、活化和存活。科学研究已经揭示了许多ECM影响细胞行为的分子信号。研究还表明，细胞生长的表面在体外确定细胞行为和优化特定细胞类型的培养条件方面起着关键作用。使用多种方法来修饰用于体外细胞培养的表面。在一种方法中，通过非特异性吸附沉积来自哺乳动物ECM的一种或多种蛋白质，例如胶原蛋白。这种涂层提供了相对先进的功能，但ECM蛋白纯化昂贵，它们必须冷藏，它们是从动物来源制备的，并且它们本身在体外控制细胞行为的能力有限。更复杂的产品如MatrigelTM可用于某些目的，但它们在分子组成方面定义不好，不适合于许多目的。合成生物基质的功能是在不使用动物产品的情况下以分子组成高度限定的方式模拟ECM的一些特征。所提出的研究的多肽多层纳米膜比现有的用于体外细胞培养的涂层具有显著的优势。这些纳米膜是完全合成的，它们可以模拟ECM蛋白的关键特征，并且它们可以针对特定的细胞类型和培养目标进行定制，这对于控制体外细胞行为的某些方面很重要。此外，这些纳米膜可能在室温下长时间储存稳定，并且与市场上现有的产品相比具有成本竞争力。拟议的研究有两个主要部分：优化一个可靠的多肽纳米膜制造过程中的多孔板，并确定不同的物理性质和生物功能化的影响，这些纳米膜上的增殖，附着和形态的各种细胞类型。如果多肽多层纳米膜显示出相对于目前市场上的涂层产品能够可靠地控制细胞附着和增殖，那么将多肽多层纳米膜技术应用于合成生物基质开发和商业化的可行性将得到证明。项目叙述哺乳动物细胞的体外培养已经成为医学研究、诊断、药物筛选、治疗性蛋白质的大规模生产以及涉及医疗保健的其他目的;对细胞培养方法的开发的兴趣持续快速增长。多肽多层纳米膜在合成生物基质材料或人工细胞外基质的开发中具有广阔的应用前景。这种材料提供了对体外细胞性质的相当或上级控制、在室温下延长的保质期和具有成本竞争力的制造，并且它们可以推进健康和疾病中的生物过程的知识，并促进新的蛋白质和细胞治疗剂的开发。

DESCRIPTION (provided by applicant): This SBIR Phase I project will determine the feasibility of applying polypeptide multilayer nanofilm technology to synthetic biomatrix development and commercialization. These nanofilms are a type of surface coating for cell and tissue culture. Immediate in vitro uses of synthetic biomatrix materials include many areas of basic medical research, diagnostic testing, high-throughput drug screening, cell phenotype selection, biologics production, and development of various therapeutic cell technologies. The synthetic biomatrix market has an estimated annual value of close to $1 billion worldwide. Any part of a tissue that is not a cell is called the extracellular matrix (ECM). In the living body the ECM surrounds cells and influences cell shape, proliferation, activation, and survival. Scientific research has revealed numerous molecular signals whereby the ECM affects cell behavior. Research has also shown that the surface on which cells are grown plays a key role in determining cell behavior in vitro and in optimizing culture conditions for a particular cell type. A variety of methods are used to modify surfaces for in vitro cell culture. In one approach, one or more proteins from mammalian ECM, for example collagen, are deposited by non-specific adsorption. Such coatings offer relatively advanced functionality, but ECM proteins are expensive to purify, they must be refrigerated, they are prepared from an animal source, and they have a limited ability on their own to control cell behavior in vitro. More complex products such as MatrigelTM are useful for some purposes, but they are poorly defined with regard to molecular composition and unsuited for many purpose. The function of a synthetic biomatrix is to mimic some features of the ECM without the use of animal products and in a way that is highly defined with regard to molecular composition. The polypeptide multilayer nanofilms of the propose research offer significant advantages over existing coatings for in vitro cell culture. These nanofilms are entirely synthetic, they can mimic crucial features of ECM proteins, and they can be tailored to a particular cell type and culture objective, important for control of some aspects of cell behavior in vitro. Moreover, these nanofilms are likely to be shelf-stable at room temperature for extended periods and cost competitive with existing products in the marketplace. The proposed research has two main parts: optimization of a reliable process of polypeptide nanofilm fabrication in multi-well plates, and determination of the effect of different physical properties and biofunctionalization of these nanofilms on the proliferation, attachment, and morphology of various cell types. Feasibility of applying polypeptide multilayer nanofilm technology to synthetic biomatrix development and commercialization will have been demonstrated if polypeptide multilayer nanofilms are shown to enable reliable control over cell attachment and proliferation relative to coating products currently on the market.Project Narrative In vitro culture of mammalian cells has become indispensable for medical research, diagnostics, drug screening, large-scale production of therapeutic proteins, and other purposes touching on healthcare; interest in development of cell culture methods continues to grow rapidly. Polypeptide multilayer nanofilms are promising for the development of synthetic biomatrix materials, or artificial extracellular matrices. Such materials offer comparable or superior control over cell properties in vitro, extended shelf-life at room temperature, and cost-competitive manufacture, and they can advance knowledge of biological processes in health and disease and promote the development of new protein and cellular therapeutics.