High Dye-Loaded and Thin-Shell Fluorescent Polymeric Nanoparticles for Enhanced FRET Imaging of Protein-Specific Sialylation on the Cell Surface

Abstract

Nanoparticle-based probes have great potential for imaging specific biomolecules in signal distinguishing and amplification via Forster resonance energy transfer (FRET). Protein-specific sialylation plays key roles in the regulation of protein structure and function, as well as in various pathophysiological processes. Here, we developed a fluorescent polymeric nanoparticle with a biocompatible hydrophilic thin shell loaded with plentiful dye and used it as the donor to enhance the FRET imaging of cell surface proteinspecific sialylation. The hydrophobic core decreased the self-quenching of loaded fluorescent molecules, while the hydrophilic thin shell ensured that the nanoparticles remained on the extracellular surface and guaranteed the FRET effect. Thus, the thin-shell polymeric nanoparticles enhanced the FRET imaging of protein tyrosine kinase-7-specific sialylation on the CCRF-CEM cell surface and showed high sensitivity under drug treatment. This nanoparticle has great potential for elucidating the relationship between dynamic specific glycosylation states and disease processes, as well as for the study of new cell surface imaging methodologies.

Softness Meets with Brightness: Dye‐Doped Multifunctional Fluorescent Polymer Particles via Microfluidics for Labeling

“…The encapsulation of cyanine dyes inside the polymer matrix is very efficient for fluorescencebased applications ranging from marker to Förster resonance energy transfer (FRET). [143][144][145] Similarly, ultrabright fluorescent polymer nanoparticles can be prepared by using rhodamine and BODIPY fluorophores. [146][147][148] Overall, specific types of dyes or their combinations can be incorporated during the formation of spherical fluorescent polymer nanoparticles of tunable sizes.…”

Nonradiative Energy Migration in Spherical Nanoparticles: Theoretical Model and Monte Carlo Study

“…Electronic excitation energy transport by the dipole–dipole Förster mechanism is a very important physical process taking place in the nanoscale up to 10 nm. Therefore, it is an effective tool to tailor, study, and describe many special systems such as nanoparticles, quantum dots, plasmonic platforms, aggregated systems, colloids, or specific ordered systems. − It can also provide relevant information about the characteristics of biologically important systems, such as end-to-end protein distance, diffusion, and conformations of fragments of macromolecules, DNA, enzymes, or albumins and serve as an efficient tool in biosensor design. − In many cases, Förster resonance energy transfer (FRET) occurs in a single step from an excited donor to an unexcited acceptor. Such a situation is, for example, typical of a macromolecule labeled by a single donor and single acceptor or more generally for the concentration of donors much lower than that of acceptors.…”

Förster Energy Transfer in Core–Shell Nanoparticles: Theoretical Model and Monte Carlo Study

“…Electronic excitation energy transfer by the dipole–dipole or Förster mechanism is an important physical process occurring in the nanoscale that is employed as an effective tool to study the properties of nanomaterials and special systems such as interfaces, fractal structures, colloids, aggregated systems, plasmonic platforms, or some ordered systems. − It is also widely used to study end-to-end distance and conformations of many macromolecules and proximity, interactions, and actions of biologically important species such as albumins, DNA, or enzymes, to name just a few. − …”