title: Site-specific peptide and protein immobilization on surface plasmon resonance chips via strain-promoted cycloaddition.
abstract: Surface plasmon resonance (SPR) is a powerful label-free diagnostic tool to study biomolecular interactions. However, one of the drawbacks of SPR is the lack of controlled immobilization of ligands on the sensor surface. We have developed a modular platform for the fast, reagent-free and site-specific immobilization of azide-containing ligands by strain-promoted cycloaddition onto a cyclooctyne-modified SPR sensor surface. The usefulness of the concept was shown in a study with a papain model system, and up to 150 experiments were performed without loss of surface quality. Furthermore, azide-containing green fluorescent protein (GFP) was also effectively immobilized. Taken together, cyclooctyne-modified SPR chips enable smooth and site-selective immobilization of ligands and prove to be more robust than traditionally functionalized systems.
authors.affiliation: Radboud University, Institute for Molecules and Materials, Nijmegen, The Netherlands.
authors.forename: Angelique E M
authors.initials: AE
authors.lastname: Wammes
authors.forename: Marcel J E
authors.initials: MJ
authors.lastname: Fischer
authors.forename: Nico J
authors.initials: NJ
authors.lastname: de Mol
authors.forename: Mark B
authors.initials: MB
authors.lastname: van Eldijk
authors.forename: Floris P J T
authors.initials: FP
authors.lastname: Rutjes
authors.forename: Jan C M
authors.initials: JC
authors.lastname: van Hest
authors.forename: Floris L
authors.initials: FL
authors.lastname: van Delft
mesh_terms: D001386:Azides / Q000737:chemistry; D061565:Cycloaddition Reaction; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D009589:Nitrogen Oxides / Q000737:chemistry; D010206:Papain / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000295:instrumentation*; D013499:Surface Properties
chemical_list: D001386:Azides; D055767:Immobilized Proteins; D009589:Nitrogen Oxides; D010455:Peptides; C015388:nitrones; D049452:Green Fluorescent Proteins; D010206:Papain