β-Particle emitting radionuclides,
such as 3H, 14C, 32P, 33P, and 35S, are
important molecular labels due to their small size and the prevalence
of these atoms in biomolecules but are challenging to selectively
detect and quantify within aqueous biological samples and systems.
Here, we present a core–shell nanoparticle-based scintillation
proximity assay platform (nanoSPA) for the separation-free, selective
detection of radiolabeled analytes. nanoSPA is prepared by incorporating
scintillant fluorophores into polystyrene core particles and encapsulating
the scintillant-doped cores within functionalized silica shells. The
functionalized surface enables covalent attachment of specific binding
moieties such as small molecules, proteins, or DNA that can be used
for analyte-specific detection. nanoSPA was demonstrated for detection
of 3H-labeled analytes, the most difficult biologically
relevant β-emitter to measure due to the low energy β-particle
emission, using three model assays that represent covalent and noncovalent
binding systems that necessitate selectivity over competing 3H-labeled species. In each model, nmol quantities of target were
detected directly in aqueous solution without separation from unbound 3H-labeled analyte. The nanoSPA platform facilitated measurement
of 3H-labeled analytes directly in bulk aqueous samples
without surfactants or other agents used to aid particle dispersal.
Selectivity for bound 3H-analytes over unbound 3H analytes was enhanced up to 30-fold when the labeled species was
covalently bound to nanoSPA, and 4- and 8-fold for two noncovalent
binding assays using nanoSPA. The small size and enhanced selectivity
of nanoSPA should enable new applications compared to the commonly
used microSPA platform, including the potential for separation-free,
analyte-specific cellular or intracellular detection.