3 research outputs found
Dynamics of Dental Pellicle Formation - <i>In Vitro</i> Analysis of Time Dependant Binding Behavior by Surface Plasmon Resonance and the Influence of Oral Therapeutics
A Novel Method for the Assessment of Targeted PEI-Based Nanoparticle Binding Based on a Static Surface Plasmon Resonance System
The delivery of nucleic acids is a major hurdle in gene therapy
or therapeutic gene knockdown, and the development of intelligent
and safe nanoparticles as carrier systems is thus under intense investigation.
The introduction of ligands for their targeted delivery is of major
interest. Here, we describe a novel approach for the analysis of the
binding properties of antibody-functionalized nanoparticles, using
surface plasmon resonance (SPR) in a static cuvette system. By chemical
coupling of the Epidermal Growth Factor Receptor (EGFR)-specific antibody
cetuximab to polyÂ(ethylene imine) (PEI) via a PEG-spacer and subsequent
DNA or siRNA complexation, we generated targeted nanoplexes with low
surface charge. Antibody-mediated uptake into EGFR overexpressing
cells was observed. SPR measurements with use of a novel, protein
A-based sandwich system for the immobilization of the target receptor
in its correct steric orientation allowed the analysis of the specific
PEI-PEG-cetuximab binding to EGFR and the determination of binding
affinities. Importantly, our cuvette-based SPR assay system was also
suitable for the monitoring of ligand-mediated nanoparticle binding,
without convection or shear stress. We conclude that our SPR sandwich
system allows the precise analysis of the binding of ligand-functionalized
nanoparticles in real-time, and we thus establish SPR for the in vitro
evaluation of ligand modifications for generating targeted nanoparticles