title: The effects of lipids and surfactants on TLR5-proteoliposome functionality for flagellin detection using surface plasmon resonance biosensing.
abstract: The use of proteoliposomes as affinity elements in conjunction with a surface plasmon resonance sensor is a high-sensitivity alternative for the detection of multiple analytes. However, one of the most important aspects of these conformations is maintaining the functionality of the immobilized protein, which is determined by the choice of lipids and surfactants employed in the reconstitutions. Previously, we demonstrated the functionality of TLR5-proteoliposomes as screening affinity elements of bacterial flagellin. In this new study we change the conditions of immobilization of TLR5 and evaluate how the fluidity of the membrane and the final size of the liposomes affect the functionality of the construct and thus increase their utility as an affinity element for design of new biosensors. In particular, we used reconstructions into preformed liposomes composed of the lipids POPC, POPC-DMPC and POPC-POPE mediated by the use of surfactants OG, Triton X100, and DDM, respectively. The affinity results were evaluated by SPR technology proteoliposomes and were correlated with the anisotropic change in the membrane status; the final sizes of the proteoliposomes were estimated. Our results clearly show the dependence of fluidity and final size of the proteoliposomes with surface plasmon resonance affinity measurements.
authors.affiliation: Biotechnology Center, Federico Santa Maria Technical University, Valparaíso, Chile. Electronic address: yusser.olguin@postgrado.usm.cl.
authors.forename: Y
authors.initials: Y
authors.lastname: Olguín
authors.affiliation: Nanobiosensor and Bioanalytical Applications Group, Institut Catàla de Nanociencia i Nanotecnología (ICN2), CSIC and CIBER-BBN, Bellaterra, Barcelona, Spain.
authors.forename: L G
authors.initials: LG
authors.lastname: Carrascosa
authors.affiliation: Nanobiosensor and Bioanalytical Applications Group, Institut Catàla de Nanociencia i Nanotecnología (ICN2), CSIC and CIBER-BBN, Bellaterra, Barcelona, Spain.
authors.forename: L M
authors.initials: LM
authors.lastname: Lechuga
authors.affiliation: Biotechnology Center, Federico Santa Maria Technical University, Valparaíso, Chile.
authors.forename: M
authors.initials: M
authors.lastname: Young
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000302:isolation & purification / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D005408:Flagellin / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D005454:Fluorescence Polarization; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D008560:Membrane Fluidity; D017830:Octoxynol / Q000737:chemistry / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011510:Proteolipids / Q000737:chemistry* / Q000378:metabolism; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance / Q000379:methods*; D013501:Surface-Active Agents / Q000737:chemistry* / Q000378:metabolism; D013997:Time Factors; D051198:Toll-Like Receptor 5 / Q000737:chemistry* / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D055767:Immobilized Proteins; D008051:Lipid Bilayers; D008055:Lipids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011510:Proteolipids; D013501:Surface-Active Agents; D051198:Toll-Like Receptor 5; C020485:proteoliposomes; C057561:1-palmitoyl-2-oleoylphosphatidylethanolamine; D005408:Flagellin; D017830:Octoxynol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; D004134:Dimyristoylphosphatidylcholine
keywords: Flagellin; Proteoliposomes; Surface plasmon resonance; Toll-like receptor 5