Ultra-sensitive detection of tumor necrosis factor alpha based on silver-coated gold core shell and magnetically separated recognition of SERS aptamer sensors.

A highly sensitive tumor necrosis factor α (TNF-α) detection method based on a surface-enhanced Raman scattering (SERS) magnetic patch sensor is reported. Magnetic beads (MNPs) and core shells were used as the capture matrix and signaling probe, respectively. For this purpose, antibodies were immobilized on the surface of magnetic beads, and then Au@4-MBN@Ag core-shell structures coupled with aptamers and TNF-α antigen were added sequentially to form a sandwich immune complex. Quantitative analysis was performed by monitoring changes in the characteristic SERS signal intensity of the Raman reporter molecule 4-MBN. The results showed that the limit of detection (LOD) of the proposed method was 4.37 × 10-15 mg·mL-1 with good linearity (R2 = 0.9918) over the concentration range 10-12 to 10-5 mg·mL-1. Excellent assay accuracy was also demonstrated, with recoveries in the range 102% to 114%. Since all reactions occur in solution and are separated by magnetic adsorption of magnetic beads, this SERS-based immunoassay technique solves the kinetic problems of limited diffusion and difficult separation on solid substrates. The method is therefore expected to be a good clinical tool for the diagnosis of the inflammatory biomarker THF-α and in vivo inflammation screening.

Self-calibration SERS sensor with "core-satellite" structure for detection of hyaluronidase activity.

Hyaluronidase expression is known to be upregulated in various pathological conditions. A method based on a combination of ratiometric surface-enhanced Raman scattering (SERS) and magnetic separation is described for the determination of hyaluronidase (HAase) activity. Gold nanospheres (AuNPs) functionalized by 4-mercaptophenylboronic acid (4-MPBA) form stable cyclic esters with diol on hyaluronic acid (HA) by the boronic acid group, while Fe3O4@DTNB@Au modified with mercaptoethylamine (MEA) was used as a capture substrate to bind to the carboxyl group on the surface of HA, forming the "Au@4-MPBA@HA/Fe3O4@DTNB@Au@MEA" "core-satellite" structure. When HAase is present, HA is enzymatically disrupted, resulting in the destruction of the "core-satellite" structure, the SERS signal of 4-MPBA is subsequently weakened. The gold shell in the substrate protects the 5,5'-Dithio bis-(2-nitrobenzoic acid) (DTNB) from the external environment, which makes it become an ideal internal standard (IS) molecule for subsequent calibration. Under optimal conditions, the I1075/I1324 varied in the range of 10-3 - 10 U‧mL-1 HAase activity, with a limit of detection (LOD) of 0.32 mU‧mL-1，below the level of HAase in normal human body fluids. This method has been successfully applied to the determination of HAase activity in urine and is expected to provide a new method in disease detection, especially in the non-invasive detection of bladder cancer.

Fabrication of Fe3O4/Au@ATP@Ag Nanorod sandwich structure for sensitive SERS quantitative detection of histamine.

We have successfully prepared a highly sensitive sandwich nanosensor combined Fe3O4 and Au@ATP@Ag nanorods for histamine detection based on surface-enhanced Raman spectroscopy (SERS). The Fe3O4 beads with -COOH served as a capture part to enrich histamine. The Au@ATP@Ag core-shell nanorods functionalized with Nalpha,Nalpha-Bis(carboxymethyl)-l-lysine (AB-NTA) were then used to connect with the imidazolyl group of histamine, simultaneously the internal standard 4-aminothiophenol (4-ATP) in the core-shell structure was used as the SERS signal. PLS regression model based on concentration range 10-3-10-8mol/L showed a linear trend with R2 = 0.9907. Our new approach can quickly and reliably determine histamine in fish sample and RAW264.7 cell lysates. This protocol for histamine extraction and SERS analysis enables the development of ultra-sensitive method for histamine detection.

Superhydrophobic silver film as a SERS substrate for the detection of uric acid and creatinine.

Superhydrophobic silver films were fabricated by silver-mirror reaction and surface functionalization with thiol. The thiol-functionalization significantly improved the hydrophobic property of the Ag films (AFS), and their contact angle values slightly increased with the extension of a thiol alkyl chain, reaching about 160°. The surface-enhanced Raman scattering (SERS) detection capacity of these films were investigated, and AFS-Dodec showed the best substrate for R6G molecule detection with the concentration limit of 10-11 M. AFS functionalized with dodecanethiol (AFS-Dodec) was applied for the SERS detection of uric acid and creatinine, it exhibited good linear dependence relationship between the Raman intensity and analyte concentration in the concentration range of 5~1000 µM.