journal article·2024Open access

A generic approach based on long-lifetime fluorophores for the assessment of protein binding to polymer nanoparticles by fluorescence anisotropy

Marwa A. Ahmed, Dóra Hessz, Benjámin Gyarmati, Mirkó Páncsics, Norbert Kovács, Róbert E. Gyurcsányi, Miklós Kubinyi, Viola Horváth

Nanoscale· Volume 16· Issue 7

Abstract

Quantitation of protein-nanoparticle interactions is essential for the investigation of the protein corona around NPs in vivo and when using synthetic polymer nanoparticles as affinity reagents for selective protein recognition in vitro. Here, a method based on steady-state fluorescence anisotropy measurement is presented as a novel, separation-free tool for the assessment of protein-nanoparticle interactions. For this purpose, a long-lifetime luminescent Ru-complex is used for protein labelling, which exhibits low anisotropy when conjugated to the protein but displays high anisotropy when the proteins are bound to the much larger polymer nanoparticles. As a proof of concept, the interaction of lysozyme with poly(N-isopropylacrylamide-co-N-tert-butylacrylamide-co-acrylic acid) nanoparticles is studied, and fluorescence anisotropy measurements are used to establish the binding kinetics, binding isotherm and a competitive binding assay.
Source: europepmc · Quality: candidate unverified · Source record ↗

Research context

OpenAlex topics: Nanoparticle-Based Drug Delivery · Protein Interaction Studies and Fluorescence Analysis · Polymer Surface Interaction Studies

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Department of Inorganic and Analytical Chemistry, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3., H-1111 Budapest, Hungary

Department of Chemistry, Faculty of Science, Arish University, 45511 El-Arish, North Sinai, Dahyet El Salam, Egypt

Department of Physical Chemistry and Materials Science, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and Economics, Műegyetem rkp. 3., H-1111 Budapest, Hungary

MTA-BME “Lendület” Quantum Chemistry Research Group, Műegyetem rkp. 3., H-1111 Budapest, Hungary

MTA-BME “Lendület” Chemical Nanosensors Research Group, Műegyetem rkp. 3., H-1111 Budapest, Hungary

ELKH-BME Computation Driven Chemistry Research Group, Műegyetem rkp. 3., H-1111 Budapest, Hungary

Funding

Magyar Tudományos Akadémia · National Research, Development and Innovation Office · Nemzeti Kutatási, Fejlesztési és Innovaciós Alap

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