Synthesis of luminescent core–shell polymer particles carrying amino groups for covalent immobilization of enzymes

Sasaki, Yusuke; Konishi, Naho; Kohri, Michinari; Taniguchi, Tadatsugu; Kishikawa, Keiki; Karatsu, Takashi

doi:10.1007/s00396-021-04913-7

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“…TOPO, originally employed in dissociation-enhanced lanthanide fluorescence immunoassays [DELFIA ® ( PerkinElmer, 2023 )], still seems to be the ancillary ligand of choice for the incorporation into beads as well ( Ando and Kawaguchi, 2005 ; Aikawa et al, 2016 ), the majority of beads being based on polystyrene, although pure PMMA is also of interest ( Moudam et al, 2009 ; Li et al, 2013 ; Cardoso Dos Santos et al, 2019 ). Last but not least, to act as biolabels, the surface of the beads has to exhibit functional groups that can readily be conjugated to the analyte under question; numerous protocols for the conjugation have been reported, protruding carboxylates and amines being the most prominent functional groups ( Petri et al, 2004 ; Hermanson and Hermanson, 2013 ; Sapsford et al, 2013 ; Sasaki et al, 2022 ): Depending on the specific method of preparation of the beads—especially the choice of the catalyst in the radical microemulsion polymerization—the beads “naturally” assume a high surface charge already (>+30 mV for 2,2′-Azobis(2-amidinopropane) dihydrochloride, AAPH, and of ca., −40 mV for Potassium peroxodisulfate, KPS). This surfcace charge is responsible for the good dispersion stability of the beads in water.…”

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confidence: 99%

High brightness red emitting polymer beads for immunoassays: Comparison between trifluoroacetylacetonates of Europium

et al. 2023

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Efficiently luminescing spherical polymer particles (beads) in the nanoscale regime of up to approximately 250 nm have become very valuable tools in bioanalytical assays. Eu3+- complexes imbedded in polymethacrylate and polystyrene in particular proved to be extraordinarily useful in sensitive immunochemical and multi-analyte assays, and histo- and cytochemistry. Their obvious advantages derive from both, the possibility to realize very high ratios of emitter complexes to target molecules, and the intrinsically long decay times of the Eu3+-complexes, which allows an almost complete discrimination against bothersome autofluorescence via time-gated measuring techniques; the narrow line emission in conjunction with large apparent Stokes shifts are additional benefits with regard to spectral separation of excitation and emission with optical filters. Last but not least, a reasonable strategy to couple the beads to the analytes is mandatory. We have thus screened a variety of complexes and ancillary ligands; the four most promising candidates evaluated and compared to each other were β-diketonates (trifluoroacetylacetonates, R-CO-CH-CO-CF3, R = - thienyl, -phenyl, -naphthyl and -phenanthryl); highest solubilities in polystyrene were obtained with trioctylphosphine co-ligands. All beads had overall quantum yields in excess of 80% as dried powders and lifetimes well beyond 600 µs. Core-shell particles were devised for the conjugation to model proteins (Avidine, Neutravidine). Their applicability was tested in biotinylated titer plates using time gated measurements and a Lateral Flow Assay as practical examples.

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