Self-assembly of robust gold nanoparticle monolayer architectures for quantitative protein interaction analysis by LSPR spectroscopy
Autor(en): | Flesch, Julia Kappen, Marie Drees, Christoph You, Changjiang Piehler, Jacob |
Stichwörter: | Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry; Chemistry, Analytical; COPOLYMERS; DYNAMICS; HOT-SPOTS; Kinetics; Localized surface plasmon resonance (LSPR); Protein immobilization; Quantitative interaction analysis; REAL-TIME; Real-time biosensor; RECEPTOR INTERACTIONS; REVEALS; Self-assembly; SERS; SINGLE-MOLECULE; TAGGED PROTEINS; TRACKING | Erscheinungsdatum: | 2020 | Herausgeber: | SPRINGER HEIDELBERG | Journal: | ANALYTICAL AND BIOANALYTICAL CHEMISTRY | Volumen: | 412 | Ausgabe: | 14, SI | Startseite: | 3413 | Seitenende: | 3422 | Zusammenfassung: | Localized surface plasmon resonance (LSPR) detection offers highly sensitive label-free detection of biomolecular interactions. Simple and robust surface architectures compatible with real-time detection in a flow-through system are required for broad application in quantitative interaction analysis. Here, we established self-assembly of a functionalized gold nanoparticle (AuNP) monolayer on a glass substrate for stable, yet reversible immobilization of Histidine-tagged proteins. To this end, one-step coating of glass substrates with poly-L-lysine graft poly(ethylene glycol) functionalized with ortho-pyridyl disulfide (PLL-PEG-OPSS) was employed as a reactive, yet biocompatible monolayer to self-assemble AuNP into a LSPR active monolayer. Site-specific, reversible immobilization of His-tagged proteins was accomplished by coating the AuNP monolayer with tris-nitrilotriacetic acid (trisNTA) PEG disulfide. LSPR spectroscopy detection of protein binding on these biocompatible functionalized AuNP monolayers confirms high stability under various harsh analytical conditions. These features were successfully employed to demonstrate unbiased kinetic analysis of cytokine-receptor interactions. |
ISSN: | 16182642 | DOI: | 10.1007/s00216-020-02551-6 |
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geprüft am 03.05.2024