Real-Time Binding Kinetics Monitored with Surface Plasmon Resonance Imaging in a Diffusion-Free Environment

R. D'Agata, G. Grasso, G. Spoto
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引用次数: 16

Abstract

Surface Plasmon Resonance Imaging (SPRI) is an optical technique emerged as a powerful tool for the simul- taneous monitoring of interactions of biomolecules arrayed onto gold substrates. To take fully advantage of the SPRI ap- proach a precise control of the fluidic of the analyte solution is imposed. The diffusion between the flowing buffer and the analyte solution which is established within the fluidic system creates a liquid volume where a gradient in the analyte con- centration is present. Such gradient is shown to affect the kinetics parameters obtained from the SPRI response. We pre- sent results obtained from a new delivery approach based on the use of air bubbles. The advantages offered by the new approach are demonstrated by using two different interacting biomolecular systems: the streptavidin-biotin system and the Ricinus Communis Agglutinin lectin-asialofetuin.
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在无扩散环境下用表面等离子体共振成像监测实时结合动力学
表面等离子体共振成像(SPRI)是一种光学技术,是同时监测排列在金底物上的生物分子相互作用的有力工具。为了充分利用SPRI方法,必须对分析物溶液的流体进行精确控制。流动缓冲液和分析物溶液之间的扩散在流体系统内建立,在分析物浓度存在梯度的情况下产生液体体积。这种梯度被证明会影响从SPRI响应中获得的动力学参数。我们介绍了一种基于气泡的新型输送方法所获得的结果。通过使用两种不同的相互作用的生物分子系统:链亲和素-生物素系统和蓖麻凝集素-凝集素-asialofetuin,证明了新方法的优势。
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