Quantitative Real-Time Imaging of Protein-Protein Interactions by LSPR Detection with Micropatterned Gold Nanoparticles
DC Element | Wert | Sprache |
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dc.contributor.author | Bhagawati, Maniraj | |
dc.contributor.author | You, Changjiang | |
dc.contributor.author | Piehler, Jacob | |
dc.date.accessioned | 2021-12-23T15:58:40Z | - |
dc.date.available | 2021-12-23T15:58:40Z | - |
dc.date.issued | 2013 | |
dc.identifier.issn | 00032700 | |
dc.identifier.uri | https://osnascholar.ub.uni-osnabrueck.de/handle/unios/3500 | - |
dc.description.abstract | Localized surface plasmon resonance (LSPR) offers powerful means for sensitive label-free detection of protein-protein interactions in a highly multiplexed format. We have here established self-assembly and surface modification of plasmonic nanostructures on solid support suitable for quantitative protein-protein interaction analysis by spectroscopic and microscopic LSPR detection. These architectures were obtained by layer-by-layer assembly via electrostatic attraction. Gold nanoparticles (AuNP) were adsorbed on a biocompatible amine-terminated poly(ethylene glycol) (PEG) polymer brush and further functionalized by poly-L-lysine graft PEG (PLL-PEG) copolymers. Stable yet reversible protein immobilization was achieved via tris(nitrilotriacetic acid) groups incorporated into the PLL-PEG coating. Thus, site-specific immobilization of His-tagged proteins via complexed Ni(II) ions was achieved. Functional protein immobilization on the surface was confirmed by real-time detection of LSPR scattering by reflectance spectroscopy. Association and dissociation rate constants obtained for a reversible protein-protein interaction were in good agreement with the data obtained by other surface-sensitive detection techniques. For spatially resolved detection, AuNP were assembled into micropatterns by means of photolithographic uncaging of surface amines. LSPR imaging of reversible protein-protein interactions was possible in a conventional wide field microscope, yielding detection limits of similar to 30 protein molecules within a diffraction-limited surface area. | |
dc.description.sponsorship | DFGGerman Research Foundation (DFG)European Commission [PI 405-4]; BMBFFederal Ministry of Education & Research (BMBF) [0312034]; We thank Gabriele Hikade and Hella Kenneweg for protein production and NB-Technologies (Bremen/Germany) for providing photomasks. This project was supported by funding from the DFG (PI 405-4) and from the BMBF (0312034). | |
dc.language.iso | en | |
dc.publisher | AMER CHEMICAL SOC | |
dc.relation.ispartof | ANALYTICAL CHEMISTRY | |
dc.subject | AFFINITY | |
dc.subject | BINDING | |
dc.subject | Chemistry | |
dc.subject | Chemistry, Analytical | |
dc.subject | FUNCTIONAL IMMOBILIZATION | |
dc.subject | HISTIDINE-TAGGED PROTEINS | |
dc.subject | LABEL-FREE DETECTION | |
dc.subject | LAYERS | |
dc.subject | MASS-TRANSPORT | |
dc.subject | POLY(ETHYLENE GLYCOL) | |
dc.subject | RECEPTOR INTERACTIONS | |
dc.subject | SURFACE-PLASMON RESONANCE | |
dc.title | Quantitative Real-Time Imaging of Protein-Protein Interactions by LSPR Detection with Micropatterned Gold Nanoparticles | |
dc.type | journal article | |
dc.identifier.doi | 10.1021/ac401673e | |
dc.identifier.isi | ISI:000326126600022 | |
dc.description.volume | 85 | |
dc.description.issue | 20 | |
dc.description.startpage | 9564 | |
dc.description.endpage | 9571 | |
dc.contributor.orcid | 0000-0002-7839-6397 | |
dc.contributor.researcherid | L-3901-2014 | |
dc.identifier.eissn | 15206882 | |
dc.publisher.place | 1155 16TH ST, NW, WASHINGTON, DC 20036 USA | |
dcterms.isPartOf.abbreviation | Anal. Chem. | |
crisitem.author.dept | Sonderforschungsbereich 944: Physiologie und Dynamik zellulärer Mikrokompartimente | - |
crisitem.author.dept | FB 05 - Biologie/Chemie | - |
crisitem.author.deptid | organisation19 | - |
crisitem.author.deptid | fb05 | - |
crisitem.author.orcid | 0000-0002-7839-6397 | - |
crisitem.author.orcid | 0000-0002-2143-2270 | - |
crisitem.author.parentorg | FB 05 - Biologie/Chemie | - |
crisitem.author.parentorg | Universität Osnabrück | - |
crisitem.author.grandparentorg | Universität Osnabrück | - |
crisitem.author.netid | YoCh745 | - |
crisitem.author.netid | PiJa938 | - |
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geprüft am 17.05.2024