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Detection of integrins using surface enhanced raman spectroscopy

机译：使用表面增强拉曼光谱法检测整联蛋白

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摘要

Integrins are transmembrane heterodimer protein receptors that mediate adherence to both the intracellular cytoskeleton and extracellular matrix. They play a major role in cellular adhesion and the breadth of their importance in biology is only recently being understood. The ability to detect concentrations of integrins on the cell surface, spatially resolve them, and study the dynamics of their behavior would be a significant advance in this field. Ultimately, the ability to detect dynamic changes of integrins on the surface of a cell maybe possible by developing a combined device such as an atomic force microscope (AFM) and surface enhanced Raman spectroscopy (SERS) system. However, the focus of this research is to first determine if integrins can be detected using SERS. Surface enhanced Raman spectroscopy (SERS) is technique used to detect the presence of analytes at the nanomolar level or below, through detection of inelastically scattered light. This thesis discusses the detection of integrins employing SERS as the detection modality. Integrins have been detected, in solution, using two silver colloids as the enhancing surface. Two silver colloid preparation methods are compared by ease of formulation and degree of enhancement in this thesis. Citrate and hydroxylamine hydrochloride (HA-HCl) reduced silver colloids were prepared through wet chemistry,compared using UV-Vis light spectroscopy, and tested for surface enhancement using adenine (a strong SERS active molecule), and two different integrins, (alpha)V(beta)3 and (alpha)5(beta)1. Results indicated that both colloids demonstrate SERS activity for varying concentrations of adenine as compared to standard non-enhanced Raman, however, only the citrate reduced colloid showed significant enhancement effect for the integrins.
机译：整联蛋白是跨膜异二聚体蛋白受体，其介导对细胞内细胞骨架和细胞外基质的粘附。它们在细胞粘附中起主要作用，并且其重要性在生物学上的广度直到最近才被了解。检测细胞表面上整联蛋白浓度，在空间上解析它们并研究其行为动力学的能力将是该领域的重大进步。最终，通过开发诸如原子力显微镜（AFM）和表面增强拉曼光谱（SERS）系统的组合设备，检测细胞表面整联蛋白动态变化的能力可能成为可能。但是，这项研究的重点是首先确定是否可以使用SERS检测到整合素。表面增强拉曼光谱（SERS）技术用于通过检测非弹性散射光来检测纳摩尔级或以下的分析物的存在。本文讨论了以SERS为检测手段对整联蛋白的检测。使用两种银胶体作为增强表面，已在溶液中检测到整联蛋白。本文比较了两种银胶体制备方法的简便性和增强程度。通过湿化学法制备柠檬酸盐和盐酸羟胺盐酸盐（HA-HCl）还原的银胶体，与UV-Vis光谱相比，并使用腺嘌呤（一种强SERS活性分子）和两种不同的整联蛋白αV测试表面增强β3和α5β1。结果表明，与标准非增强拉曼相比，两种胶体均表现出不同浓度腺嘌呤的SERS活性，但是，只有柠檬酸盐还原的胶体对整联蛋白显示出显着的增强作用。

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    Gant Virgil Alexander;

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  • 年度 2005
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