Antibiotics pp 145-157 | Cite as

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

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1520)

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.

Key words

Biosensors Surface acoustic wave (SAW) Model membranes 

Notes

Acknowledgments

The authors would like to thank Hildegard Falkenstein-Paul for excellent technical competences in performing the SAW measurements and providing the data presented here.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Pharmaceutical Chemistry II, Pharmaceutical InstituteUniversity of BonnBonnGermany

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