Surface Plasmon Resonance Bioanalytical Platform to Appraise the Interaction Between Antimicrobial Peptides and Lipid Me
Surface Plasmon Resonance (SPR) sensors gain a wide applicability as a direct, label-free, and real-time approach to analyze biomolecular reactions occurring in the vicinity of a functionalized sensor surface. Lipid-modified sensor chips provide an access
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Abstract Surface Plasmon Resonance (SPR) sensors gain a wide applicability as a direct, label-free, and real-time approach to analyze biomolecular reactions occurring in the vicinity of a functionalized sensor surface. Lipid-modified sensor chips provide an accessible platform for SPR exploration of membrane—peptide interactions. While pore formation and subsequent membrane destabilization is a common feature for the interaction process between a large number of compounds such as peptides, toxins, and viruses with lipid membranes, this process has been just recently related to sensing applications. Using POPC and melittin as model systems we show that SPR quantitative appraisal of the interaction between an antimicrobial peptide and lipid-modified sensors is capable to provide both novel sensing avenues and detailed mechanistic insights into effects of pore-forming compounds. This new and exciting biosensing avenue is based on assessment of the nonmonotonous, concentration-dependent effect of pore formation and enables quantitative evaluation of the whole process, including full dissolution of the lipid. Insight will be provided on a novel kinetic model that relates, via the Transfer Matrix, surface plasmon resonance (SPR) data with actual concentrations of interacting partners. In agreement with literature data, association and dissociation rates, concentration thresholds, evolution within each interacting layer of lipid and peptide concentrations, as well as of peptide to lipid ratios are derived. Although based on Biacore 3000 data, the general principles and guidelines may be applicable to other SPR assays. This biosensing approach is suitable to an entire set of pore-forming compounds including antimicrobial peptides and toxins and different lipid matrices. Recent developments in terms of surface
M. Gheorghiu (*) • S. David • A. Olaru • C. Polonschii • E. Gheorghiu International Centre of Biodynamics, 1 B Intrarea Portocalelor, 060101 Bucharest, Romania e-mail: [email protected] http://www.biodyn.ro W. Fritzsche and J. Popp (eds.), Optical Nano- and Microsystems for Bioanalytics, Springer Series on Chemical Sensors and Biosensors (2012) 10: 183–210 DOI 10.1007/978-3-642-25498-7_6, # Springer-Verlag Berlin Heidelberg 2012
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functionalization and electro-optical integration toward a portable analytical platform are discussed. The proposed approach combined with appropriate design of the experimental protocol adds a new depth to the classic SPR investigation of peptide–lipid interaction offering a quantitative platform for detection, improved understanding of the manifold facets of the interaction, and for supporting the controlled design of novel antimicrobial compounds. Keywords Bio(mimetic) sensing platform • EIS • Kinetic analysis • SPR
Contents 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Why Antimicrobial Peptides? . . . . . . . . . . . . . .
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