A novel peptide/Fe3O4@SiO2-Au nanocomposite-based fluorescence biosensor for the highly selective and sensitive detection of prostate-specific antigen.

Highly selective and sensitive detection methods are very important for the early diagnosis of prostate-specific antigen (PSA). Here, we present a novel peptide/Fe3O4@SiO2-Au nanocomposite-based fluorescence biosensor for highly selective and sensitive detection of PSA. The biosensor was made by self-organizing 5-FAM labeled peptides onto the surface of magnetic Fe3O4@SiO2-Au nanocomposites (MNCPs), resulting in efficient quenching of the FAM fluorescence. The PSA specifically recognized and cleaved the 5-FAM-labeled peptides, leading to the fluorescence recovery. This is the first report of the MNCPs by in situ growth of Au nanoparticles (AuNPs) on the SiO2 encapsulated single Fe3O4 nanocubes. The MNCPs feature robust salt stability, and allow for effective fluorescence quenching and easy magnetic separation, which greatly decrease the background fluorescence. The peptide/MNCPs-based fluorescence biosensor measure a wide range of concentrations of PSA, from 1.0 × 10-12 to 1.0 × 10-9g/mL, with a limit of detection (LOD) of 3.0 × 10-13g/mL in both standard solutions and serum samples, demonstrating the great potential of this biosensor platform for use in clinical and biological assays.

A fluorescence aptasensor based on two-dimensional sheet metal-organic frameworks for monitoring adenosine triphosphate.

In the present study, a facile fluorescence aptasensor based on two-dimensional sheet metal-organic frameworks of N,N-bis(2-hydroxyethyl)dithiooxamidato copper(II) (H2dtoaCu) was developed for the sensitive detection of adenosine triphosphate (ATP). The sensing mechanism was based on the noncovalent interaction between FAM-labeled (fluorescein amidite) ATP aptamers and H2dtoaCu. In the absence of ATP, the FAM-labeled aptamer readily adsorbs onto H2dtoaCu, mainly via π-π stacking and hydrogen bond interactions between the nucleotide bases and the H2dtoaCu surface, leading to the reduction of fluorescence intensity of the FAM by photoinduced electron transfer (PET). In the presence of ATP, the FAM-labeled aptamer specifically forms ATP-binding aptamer complexes which exhibit only weak adsorption on the H2dtoaCu surface. Thus, the fluorescence of the FAM-labeled ATP aptamer remained largely unchanged. The fluorescence aptasensor exhibited a good linear relationship between the fluorescence intensity and the logarithm concentration of ATP over a range of 25-400 nM, with a detection limit of 8.19 nM (3S/N). ATP analogs such as guanosine triphosphate, uridine triphosphate, and cytidine triphosphate have negligible effect on the aptasensor performance due to the high selectivity of the ATP aptamer to its target, showing promising potential in real sample analysis.

Carbohydrate analysis on hybrid poly(dimethylsiloxane)/glass chips dynamically coated with ionic complementary peptide.

A facile and efficient dynamic coating method using an ionic complementary peptide was established for high-performance separation of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled carbohydrates in a hybrid poly(dimethylsiloxane) (PDMS)/glass microfluidic channel. EAK16-II with a sequence of [(Ala-Glu-Ala-Glu-Ala-Lys-Ala-Lys)2] can readily self-organize into a complete coating layer tightly adsorbed on both hydrophobic PDMS and hydrophilic glass surfaces, which efficiently suppressed nonspecific analyte adsorption and minimized electroosmotic flow (EOF). Separation conditions were systematically investigated with respect to EAK16-II concentration, running buffer, buffer pH, and field strength (Esep). Under the optimal conditions, rapid and reproducible separations of maltodextrin ladder, glycans from glucosamine capsules, tablets, and pomegranate peel extracts were achieved with over 450000 theoretical plates per meter in the hybrid PDMS/glass microchannels dynamically coated with 1.0mg/mL EAK16-II-0.05% n-dodecyl ß-d-maltoside (DDM), and the relative standard deviation (RSD) values were less than 3.2% (n=4) for the migration times. The present work provides a facile and efficient means to minimize EOF and nonspecific analyte adsorption in microfluidic chips fabricated in various substrates, thereby broadening the applications of microfluidic chips in complicated biological assays.

MnO2 nanosheet-assisted ligand-DNA interaction-based fluorescence polarization biosensor for the detection of Ag+ ions.

Silver (Ag+) ions are highly toxic to aquatic organisms and accumulate in the human body via the food chain. Therefore, the development of sensitive and selective quantitative analytical methods for detecting trace amounts of these ions is necessary. In the present study, a MnO2 nanosheet-assisted, ligand-DNA interaction and fluorescence polarization-based method was developed, for the first time, for sensitive detection of Ag+. The addition of Ag+ to the preformed proflavine-DNA complex induced the release of proflavine, which elicited weak changes in fluorescence polarization. The subsequent addition of MnO2 nanosheets magnified the observed changes, making this a feasible method for the detection of Ag+. The calibration graphs indicated good linearity over the concentration ranges of 30-240nM for Ag+, with a detection limit (S/N=3) of 9.1nM. This method additionally exhibits high selectivity. The mechanism underlying the changes in fluorescence polarization caused by the addition of Ag+ in the presence of MnO2 nanosheets was further explored in this study. These findings demonstrate that the present MnO2 nanosheet-assisted fluorescence polarization biosensor may represent a promising tool for the detection of Ag+ ions. The results for practical detecting Ag+ proved that this biosensor can be applied to environmental water sample analysis.