Holistic Assessment of Covalently Labeled Core–Shell
Polymeric Nanoparticles with Fluorescent Contrast Agents for Theranostic
Applications
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Abstract
The successful development of degradable
polymeric nanostructures
as optical probes for use in nanotheranostic applications requires
the intelligent design of materials such that their surface response,
degradation, drug delivery, and imaging properties are all optimized.
In the case of imaging, optimization must result in materials that
allow differentiation between unbound optical contrast agents and
labeled polymeric materials as they undergo degradation. In this study,
we have shown that use of traditional electrophoretic gel-plate assays
for the determination of the purity of dye-conjugated degradable nanoparticles
is limited by polymer degradation characteristics. To overcome these
limitations, we have outlined a holistic approach to evaluating dye
and peptide–polymer nanoparticle conjugation by utilizing steady-state
fluorescence, anisotropy, and emission and anisotropy lifetime decay
profiles, through which nanoparticle–dye binding can be assessed
independently of perturbations, such as those presented during the
execution of electrolyte gel-based assays. This approach has been
demonstrated to provide an overall understanding of the spectral signature–structure–function
relationship, ascertaining key information on interactions between
the fluorophore, polymer, and solvent components that have a direct
and measurable impact on the emissive properties of the optical probe.
The use of these powerful techniques provides feedback that can be
utilized to improve nanotheranostics by evaluating dye emissivity
in degradable nanotheranostic systems, which has become increasingly
important as modern platforms transition to architectures intentionally
reliant on degradation and built-in environmental responses