Synthesis of surface protein-imprinted nanoparticles based on metal coordination and anchored carbon dots for enhanced fluorescence detection.

Molecularly imprinted polymers endowed with photo-luminescent properties have attracted wide research interest in many fields such as biological analysis and diseases diagnosis. Herein, we illustrate a versatile method for the construction of surface protein-imprinted nanoparticles based on metal coordination and anchored carbon dots (CDs) for enhanced fluorescence detection of the target protein. As the fluorescent nanosupports for surface imprinting, CDs-attached SiO2 nanoparticles were synthesized via thiol-ene click chemistry. With histidine (His)-exposed protein as templates, imprinted nanoshells were formed over the nanosupports via copolymerization of a Cu2+-chelating monomer and an oligo (ethylene glycol) monomer, hence producing high-quality imprinted cavities because of both the relatively strong coordination and inhibited non-specific binding. Using lysozyme as a model His-exposed template, the imprinted nanoparticles showed fluorescence enhancement while binding the target protein, and exhibited significantly increased specific fluorescence response than the controls without the metal coordination. They achieved a high imprinting factor of 5.8 and a low limit of detection of 10.1 nM. Furthermore, such sensors were applied to determine lysozyme in diluted chicken egg-white samples with satisfactory recoveries at three spiking levels ranging from 97.9 to 101.4%. Human serum albumin was also used as another template protein for preliminary confirming the generality of the presented strategy.

A method for synthesis of oriented epitope-imprinted open-mouthed polymer nanocapsules and their use for fluorescent sensing of target protein.

Epitope imprinting has proved to be an effective way for fabricating artificial receptors for protein recognition. Surface imprinting over sacrificial supports is particularly favorable for generating high-quality epitope-imprinted cavities, but obtaining nanomaterials by this way is still a challenge. Herein, we propose a method for the synthesis of oriented surface epitope-imprinted open-mouthed polymer nanocapsules (OM-MIP NCs) by sacrificing asymmetric template-modified Janus nanocores. Amine/aldehyde functionalized SiO2 Janus nanoparticles were prepared via the molten-wax-in-water Pickering emulsion approach, an easy scale-up technique. Epitope templates and vinyl groups were coupled to the aldehyde-bearing major side, whereas polyethylene glycol (PEG) chains were grafted to the amine-modified side. Incomplete imprinted shells were then generated principally on the non-PEGylated side via aqueous precipitation polymerization, hence affording OM-MIP NCs after etching the SiO2 nanocores. With a C-terminus nonapeptide of bovine serum albumin (BSA) chosen as a model epitope and polymerizable carbon dots added to the pre-polymerization solution, fluorescent OM-MIP NCs were synthesized for sensing of BSA. Such NCs reached maximal fluorescent response within 15 min, greatly faster than the closed imprinted NCs within 130 min, proving good accessibility of their inner-surface imprinted cavities thanks to the open mouths. Furthermore, they showed excellent target protein detection performance, with an imprinting factor of 7.8, a limit of detection of 43.8 nM and a linear range of 0.2-6 µM. The recoveries in bovine serum samples at four spiking levels ranged from 99.2 to 107.2%, with relative standard deviations of 1.2-5.9%.

NADH induces iron release from pea seed ferritin: a model for interaction between coenzyme and protein components in foodstuffs.

Plant ferritin from legume seeds co-exists with coenzyme NADH (a reduced form of nicotinamide-adenine dinucleotide) in many foodstuffs. In the present study, the interaction of NADH with apo pea seed ferritin (PSF) was investigated by fluorescence resonance energy transfer (FRET), fluorescence titration, transmission electron microscope (TEM), and isothermal titration calorimetry (ITC). We found that NADH molecules bound on the outer surface of PSF close to the 4-fold channels, which was 1.58 nm from tryptophan residue (Trp). Consequently, such binding facilitates iron release from holo PSF, which might have a negative effect on PSF as an iron supplement, while NADH was oxidised into NAD(+). However, the binding of NADH to the protein does not affect the entry of toxic ferrous ions into the apo protein shell, where these ferrous ions were oxidised into less toxic ferric ions. Moreover, NADH binding markedly affects the tertiary structure around Trp residues of PSF. These findings advanced our understanding of the interactions between different naturally occurring components in a complex food system.