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用于确定分析物膜相互作用的质量敏感生物传感器系统。

Mass-Sensitive Biosensor Systems to Determine the Membrane Interaction of Analytes.

作者信息

Hoß Sebastian G, Bendas Gerd

机构信息

Pharmaceutical Chemistry II, Pharmaceutical Institute, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.

出版信息

Methods Mol Biol. 2017;1520:145-157. doi: 10.1007/978-1-4939-6634-9_9.

Abstract

Biosensors are devices that transform a biological interaction into a readout signal, which is evaluable for analytical purposes. The general strength of biosensor approaches is the avoidance of time-consuming and cost-intensive labeling procedures of the analytes. In this chapter, we give insight into a mass-sensitive surface-acoustic wave (SAW) biosensor, which represents an elegant and highly sensitive method to investigate binding events at a molecular level. The principle of SAW technology is based on the piezoelectric properties of the sensors, so as to binding events and their accompanied mass increase at the sensor surface are detectable by a change in the oscillation of the surface acoustic wave. In combination with model membranes, transferred to the sensor surface, the analytical value of SAW biosensors has strongly been increased and extended to different topics of biomedical investigations, including antibiotic research. The interaction with the bacterial membrane or certain target structures therein is the essential mode of action for various antibacterial compounds. Beside targeted interaction, an unspecific membrane binding or membrane insertion of drugs can contribute to the antibacterial activity by changing the lateral order of membrane constituents or by interfering with the membrane barrier function. Those pleiotropic effects are hardly to illustrate in the bacterial systems and need a detailed view at the in vitro level. Here, we illustrate the usefulness of a SAW biosensor in combination with model membranes to investigate the mode of membrane interaction of antibiotic active peptides. Using two different peptides we exemplary describe the interaction analysis in a two-step gain of information: (1) a binding intensity or affinity by analyzing the phase changes of oscillation, and (2) mode of membrane interaction, i.e., surface binding or internalization of the peptide by following the amplitude of oscillation.

摘要

生物传感器是将生物相互作用转化为可用于分析目的的读出信号的装置。生物传感器方法的总体优势在于避免了对分析物进行耗时且成本高昂的标记程序。在本章中,我们将深入探讨一种质量敏感型表面声波(SAW)生物传感器,它是一种在分子水平上研究结合事件的精巧且高度灵敏的方法。SAW技术的原理基于传感器的压电特性,因此传感器表面的结合事件及其伴随的质量增加可通过表面声波振荡的变化来检测。与转移到传感器表面的模型膜相结合,SAW生物传感器的分析价值得到了极大提升,并扩展到生物医学研究的不同领域,包括抗生素研究。与细菌膜或其中某些靶标结构的相互作用是各种抗菌化合物的基本作用方式。除了靶向相互作用外,药物的非特异性膜结合或膜插入可通过改变膜成分的侧向排列或干扰膜屏障功能来促进抗菌活性。这些多效性效应在细菌系统中很难阐明,需要在体外水平进行详细观察。在此,我们展示了SAW生物传感器与模型膜相结合在研究抗生素活性肽膜相互作用模式方面的实用性。我们使用两种不同的肽,通过两步获取信息的方式示例性地描述相互作用分析:(1)通过分析振荡的相位变化来确定结合强度或亲和力,(2)通过跟踪振荡幅度来确定肽的膜相互作用模式,即表面结合或内化。

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