Mobility of Proteins Adsorbed at Solid-Liquid Interfaces

这项工作的主要目标是表征吸附在固-液界面上的蛋白质的流动性。这项研究代表了蛋白质吸附研究的重点转向对被吸附蛋白质的复杂行为的分子理解。人们希望量化吸附蛋白质的迁移率，表征影响迁移率的因素，确定吸附蛋白质翻译的机制，并测量蛋白质在表面岛上自组装的程度。结合使用全内反射荧光和图案光漂白后的荧光恢复，这项研究将确定蛋白质表面扩散系数和可移动的吸附蛋白质的比例。流动性将被量化为剪切率、表面覆盖率和表面停留时间的函数，以确定控制因素。了解吸附蛋白质的流动性，以及蛋白质结构和组织的相关问题，将有助于开发有效的工艺来制造蛋白质和多肽材料。这些材料的用途正在增加，不仅作为药物，而且作为有机催化的新手段，以及作为化学分析的生物传感器的活化剂。在制造、修饰或提纯这些材料的所有过程中，蛋白质或多肽在固体表面的吸附是不可避免的。这对蛋白质和多肽的制造特别重要，因为吸附的蛋白质层可以影响其接触的细胞的生物活性，包括工业上常用的基因工程微生物。此外，蛋白质或多肽的正常功能依赖于正确结构的保存。吸附到固体表面会改变这种结构。

The primary objective of this work is to characterize the mobility of proteins adsorbed at solid-liquid interfaces. This research represents a shift in the emphasis of protein adsorption studies towards a molecular understanding of the complex behavior of adsorbed proteins. It is hoped to quantify the mobility of adsorbed proteins, characterize the factors governing mobility, identify the mechanism of adsorbed protein translation, and measure the extent of self-assembly of proteins into surface islands. Using a combination of total internal reflection fluorescence and fluorescence recovery after pattern photobleaching, the research will determine protein surface diffusion coefficients and the fraction of adsorbed proteins that are mobile. Mobility will be quantified as a function of shear rate, surface coverage, and surface residence time in order to identify the governing factors. Understanding the mobility of adsorbed proteins, as well as the related issues of protein structure and organization will aid the development of effective processes to manufacture protein and polypeptide materials. The utility of these materials is increasing, not only as pharmaceuticals, but also as novel means of organic catalysis, and as the active agent in biosensors for chemical analysis. Protein or polypeptide adsorption to solid surfaces is inevitable in all processes to manufacture, modify, or purify these materials. This is particularly important to protein and polypeptide manufacture in that an adsorbed protein layer can influence the biological activity of the cells it contacts, including the genetically engineered microorganisms commonly employed in industry. In addition, the proper function of a protein or polypeptide depends on the preservation of the correct structure. Adsorption to a solid surface can alter that structure.