title: Application of Bio-Layer Interferometry for the analysis of protein/liposome interactions.
abstract: The development of biosensor technologies for the investigation of biomolecular interactions has markedly advanced over the last years. One promising biosensor platform, the Bio-Layer Interferometry (BLI), was developed by ForteBio with the main focus to qualify and quantify protein/protein interactions in research and routine applications. Here, a method to characterize protein/liposome binding interactions based on the biophysical principles of this platform is described. Three different liposome formulations and the protein hormone, recombinant human erythropoietin (rh-Epo) were used as models in the test system. Rh-Epo was immobilized on disposable optical fiber streptavidin (SA) biosensor tips and binding of different liposome formulations under certain conditions was measured. The assay performance was evaluated, followed by calculating the kinetic rate and affinity constants. The results showed that all liposome formulations formed extremely stable complexes with the immobilized protein. Nevertheless, liposome specific differences in binding affinities were determined. Furthermore, a liposome concentration dependent binding pattern was demonstrated. The combination of simple sample preparation, the opportunity of automation with high throughput in an acceptable time range and excellent reproducibility, makes this assay suitable for basic research as well as for drug discovery and drug screening to estimate drug/membrane interactions.
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Muthgasse 11, A-1190 Vienna, Austria. Jakob.Wallner@boku.ac.at
authors.forename: Jakob
authors.initials: J
authors.lastname: Wallner
authors.forename: Gabriele
authors.initials: G
authors.lastname: Lhota
authors.forename: Dominik
authors.initials: D
authors.lastname: Jeschek
authors.forename: Alexander
authors.initials: A
authors.lastname: Mader
authors.forename: Karola
authors.initials: K
authors.lastname: Vorauer-Uhl
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D055808:Drug Discovery / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004921:Erythropoietin / Q000378:metabolism; D006728:Hormones / Q000378:metabolism; D006801:Humans; D007368:Interferometry / Q000379:methods*; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism*; D055100:Optical Fibers; D011485:Protein Binding / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D019809:Streptavidin / Q000378:metabolism
chemical_list: D006728:Hormones; D008081:Liposomes; D011994:Recombinant Proteins; D004921:Erythropoietin; D019809:Streptavidin