Using the High-Entropy Approach to Obtain Multimetal Oxide Nanozymes: Library Synthesis, In Silico Structure-Activity, and Immunoassay Performance.

High-entropy nanomaterials exhibit exceptional mechanical, physical, and chemical properties, finding applications in many industries. Peroxidases are metalloenzymes that accelerate the decomposition of hydrogen peroxide. This study uses the high-entropy approach to generate multimetal oxide-based nanozymes with peroxidase-like activity and explores their application as sensors in ex vivo bioassays. A library of 81 materials was produced using a coprecipitation method for rapid synthesis of up to 100 variants in a single plate. The A and B sites of the magnetite structure, (AA')(BB'B'')2O4, were substituted with up to six different cations (Cu/Fe/Zn/Mg/Mn/Cr). Increasing the compositional complexity improved the catalytic performance; however, substitutions of single elements also caused drastic reductions in the peroxidase-like activity. A generalized linear model was developed describing the relationship between material composition and catalytic activity. Binary interactions between elements that acted synergistically or antagonistically were identified, and a single parameter, the mean interaction effect, was observed to correlate highly with catalytic activity, providing a valuable tool for the design of high-entropy-inspired nanozymes.

Investigating conjugated polymer nanoparticle formulations for lateral flow immunoassays.

Lateral flow immunoassays (LFI) are valuable tools for point-of-care testing. However, their sensitivity is limited and can be further improved. Nanoparticles (NP) of conjugated polymers (CPNs), also known as Pdots, are reported to be highly sensitive fluorescent probes, but a direct comparison with conventional colloidal gold-based (Au-NP) LFI using the same antibody-antigen pair is missing to date. Furthermore, the influence of brightness and Stokes shift of CPs on the signal : background ratio (SBR) needs to be evaluated. In this study, we encapsulated two different CPs, poly-(9,9-di-n-octyl-fluorenyl-2,7-diyl) (PDOF) and poly-(2,5-di-hexyloxy-cyanoterephthalylidene) (CN-PPV) in silica shell-crosslinked Pluronic© micelles (Si-NP) and Pdots and investigated the NP brightness with respect to CP loading dose. The brightest formulation of each NP system was conjugated to rabbit IgG as a model antigen and the SBR was investigated in an ELISA-like microplate assay and LFI. Two reference particles, Au-NP and a polystyrene NP (PS-NP) loaded with a small-molecule fluorescent dye were conjugated to IgG and compared to the Si-NP and Pdots. The mass of Pdots required for detection in LFI was at least two orders of magnitude lower than that of Si-NP and the reference NP. The SBR of CN-PPV (moderate brightness, large Stokes shift) was two to three times higher than the SBR of PDOF (high brightness, small Stokes shift). To combine the favourable properties of both CPs, a polymer blend of PDOF and CN-PPV was encapsulated in Pdots, and resulted in further increase of SBR in the microplate assay and LFI. In summary, combining two CPs with different properties can lead to fluorescent signal-transducers for applications such as ELISA and LFIs, which can enhance the detection limit of the assay by 2-3 orders of magnitude.

Enhanced optical imaging properties of lipid nanocapsules as vehicles for fluorescent conjugated polymers.

Conjugated polymer nanoparticles (CPNs) have emerged as highly photostable probes for optical and photoacoustic imaging. However, the aggregation of conjugated polymer (CP) molecules upon nanoparticle formation is associated with fluorescence quenching, poor yields and mutable particle sizes. This study investigated whether the CP encapsulation within the liquid midchain triglyceride (MCT) core of lipid nanocapsules (LNCs) may achieve reduced packing of CP chains leading to a stable system with enhanced optical features. The red- and near infrared-emitting CPs, CN-PPV and PCPDTBT, showed precipitation and aggregation-induced quenching with concentrations >~25 µg/mL in MCT alone. Despite this, CP encapsulation within LNCs abolished quenching at concentrations up to 1500 µg/mL. PCPDTBT-LNCs exhibited a quantum yield of 2.8% and a higher signal:background ratio in an optical imaging phantom compared to literature reports of PCPDTBT encapsulated in PEG-PLGA nanoparticles. In contrast, PCPDTBT-LNCs had slightly lower photoacoustic amplitudes than reported PEG-PLGA systems. CP-LNCs were also stable in size (32 ± 0.7 nm) and photoluminescence over 21 days at 4 °C, 25 °C and 37 °C. In summary, encapsulation of CP within the liquid core of lipid nanocapsules enhances the optical properties of fluorescent CP.