In this study, a simple and highly selective homogeneous sandwich assay was developed for fast and ultrasensitive detection of the tau protein using a combination of monoclonal antitau functionalized hybrid magnetic nanoparticles and polyclonal antitau immobilized gold nanoparticles as the recognition and surface-enhanced Raman scattering (SERS) component, respectively. The magnetic silica particles were first coated with poly(2-hydroxyethyl methacrylate) via surface-mediated reversible addition-fragmentation chain transfer (RAFT) polymerization and then biofunctionalized with monoclonal antitau, which are both specific for tau and can be collected via a simple magnet. After separating tau from the sample matrix, they were sandwiched with the SERS substrate composed of polyclonal antitau and 5,5-dithiobis(2-dinitrobenzoic acid) on gold nanoparticles. The correlation between the tau concentration and SERS signal was found to be linear within the range of 25 fM to 500 nM. The limit of detection for the sandwich assay is less than 25 fM. Moreover, the sandwich assay was also evaluated for investigating the tau specificity on bovine serum albumin and immunoglobulin G.
In this report, we present a new homogeneous detection method for staphylococcal enterotoxin B (SEB) utilizing core-shell-structured iron-gold magnetic nanoparticles and a gold nanorod surface-enhanced Raman scattering (SERS) probe in solution. Peptide ligand (aptamer) functionalized magnetic gold nanorod particles were used as scavengers for target SEB. After the SEB molecules were separated from the matrix, the sandwich assay procedure was tested by gold nanorod particles that act as SERS probes. The binding constant between SEB and peptide-nanoparticle complex was determined as 8.0 × 10(7) M(-1). The correlation between the SEB concentration and SERS signal was found to be linear within the range of 2.5 fM to 3.2 nM. The limit of detection for the homogeneous assay was determined as 224 aM (ca. 2697 SEB molecules/20 μL sample volume). Also, gold-coated surfaces were used as capture substrates and performances of the two methods were compared. Furthermore, the developed method was evaluated for investigating the SEB specificity on bovine serum albumin (BSA) and avidin and detecting SEB in artificially contaminated milk, blood, and urine.
A general protocol to prepare surface molecularly imprinted polymer core-shell superparamagnetic Fe3O4 nanoparticles (Fe3O4@MIP SPNPs), using a surface-mediated RAFT polymerization approach, is described. Cholesterol-imprinted Fe3O4@MIP SPNPs were obtained by oleic acid-stabilized Fe3O4 nanoparticles with a trithiocarbonate agent and subsequently by polymerizing thin molecularly imprinted layers composed of dimethylacrylamide and N,N'-methylene(bis)acrylamide units. The surface-mediated RAFT polymerization approach allows the synthesis of ∼20 nm hybrid composite particles with a ∼6 nm MIP shell, exhibiting superparamagnetic properties (saturation magnetization = 35.4 emu g(-1)) and specific molecular recognition of cholesterol. The Fe3O4@MIP SPNPs show the capability of rapid enriching and separating cholesterol (∼3.1% in weight) and are renewable and cyclically exploited due to their monodispersive and superparamagnetic features. Moreover, under optimal conditions, the Fe3O4@MIP SPNP recoveries of spiked human serum, milk, yolk and beef were 91.6%, 93.6%, 92.4% and 91.2%, respectively. Finally, the method of molecular imprinting on superparamagnetic particles can be extended to a wide range of applications for cell sorting, biomolecule enrichment and separation, and drug delivery.
In this report, we have developed a novel surface-enhanced Raman scattering (SERS) aptasensor for ricin B toxin recognition based on Ag nanoparticles labeled with 4,4 0 -bipyridyl (Bpy, Raman reporter) and ricin B aptamer. The hybrid silicon substrate was first prepared via surface-mediated RAFT polymerization of Nacryoyl-L-valine in the presence of 2-(butylthio-carbonothioylthio)-2-methylpropionic acid-modified silicon wafer as a RAFT agent and then biofunctionalized with ricin B aptamer. In this novel system, the ricin B aptamer functionalized silicon substrate was used as a scavenger for target ricin B molecules.After ricin B molecules were separated from the matrix, the sandwich assay procedure was applied using Ag nanoparticles labeled with Bpy and ricin B aptamer which act as SERS probes. Meanwhile, to enhance the SERS signal, silver deposition on the sandwich complex was also performed. The correlation between the ricin B concentration and SERS signal was found to be linear within the range of 1.0 fM to 50 pM. The limit of detection for the SERS aptasensor was determined as 0.32 fM. Furthermore, the SERS aptasensor was also evaluated for detecting ricin B in artificially contaminated orange juice, milk, blood and urine. Finally, this method has the potential to be used for the detection of other protein toxins in a complex matrix if a specific aptamer for that protein toxin can be designed.Scheme 1 Schematic representation of the synthesis of poly(AVAL) brushes on the silicon surface. Fig. 1 N1s, C1s and S2p core level XPS spectra of (a) Si-GPTS, (b) Si-BCTP and (c) poly(AVAL) brushes.
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