Phenylethanolamine A (PA) is a new kind of β-agonist, which was illegally used as a feed additive for growth promotion in China. In this study, a novel immunochromatographic assay (ICA) based on surface-enhanced Raman scattering (SERS) for the ultrasensitive and quantitative detection of phenylethanolamine A is presented. The principle of this new ICA is similar to that based on colloidal gold particles, but using Au(MBA)@Ag-Ab [e.g., polyclonal antibody of PA labeled Au@Ag core-shell nanoparticles (NPs) sandwiched with a Raman reporter (4-mercaptobenzoic acid, MBA)] as a probe. After ICA procedures, the specific Raman scattering intensity of MBA on the test line was measured for quantitative detection of PA. This assay was completed within 15 min. The IC50 and limit of detection (LOD) values of the ICA for PA detection were 0.06 ng mL(-1) and 0.32 pg mL(-1), respectively, which were 1-3 orders of magnitude lower than those obtained by other immunoassays, indicating the ultrasensitivity of this ICA. There was no cross-reactivity (CR) of the assay with another three β-agonists (ractopamine, clenbuterol, and salbutamol), suggesting high specificity of the SERS-based ICA. A spiking experiment revealed that the recoveries of PA from pig urine samples were in range of 99.9- 101.2% with relative standard deviations (RSDs) of 3.6-5.8%. The results demonstrated that this SERS-based ICA was able to quantitatively detect PA in urine samples with high sensitivity, specificity, precision, and accuracy and might be a powerful method for the analysis of other target analytes in the food area.
A multiple signal amplification strategy was designed for an ultrasensitive competitive immunoassay for Hg(2+). This strategy was achieved using graphene conjugated with a large number of CdSe quantum dots to enhance the basal signal and enormous horseradish peroxidase (HRP) labeled with gold nanoparticles (AuNPs) to consume the coreactant H2O2 generated in situ. The immunosensor was constructed by immobilization of coating antigen on poly(diallyldimethylammonium chloride)-graphene-CdSe composites (PDDA-GN-CdSe), and a strong electrochemiluminescence (ECL) signal was obtained. When the immunosensor was immersed in antibody-AuNPs-HRP composites, the ECL signal greatly decreased, which was ascribed to the bound enzyme on the electrode surface. The self-produced coreactant H2O2 was consumed by o-phenylenediamine in the presence of enzyme, effectively decreasing the ECL intensity from the quantum dots. The Hg(2+) in solution and the corresponding coating antigen competed for the limited antibody, and thus, the ECL intensity was linearly dependent on the logarithm of the mercury(II) concentration from 0.2 to 1000 ng mL(-1) with a detection limit of 0.06 ng mL(-1). The immunoassay exhibited good stability and accuracy and acceptable reproducibility, indicating that it provides a promising approach for the detection of trace mercury and other small molecular compounds in environmental samples.
The yields of NO from combustion of bituminous coal, lignite, and biomass chars were investigated in O 2 /N 2 and O 2 /CO 2 atmospheres. The experiments were performed in a laboratory-scale fixed-bed reactor in the temperature range of 850− 1150 °C. To minimize thermal deactivation during char preparation, the chars were generated by in situ pyrolysis at the reaction temperature. The NO yield clearly decreased and the CO yield increased when the atmosphere was altered from O 2 /N 2 to O 2 /CO 2 at 850 °C, but only small differences in NO and CO yields were observed between the two atmospheres at 1050−1150 °C. To examine how CO influences the NO yield, the effect of CO on NO reduction over char as well as NO reduction by CO over ash was investigated in the fixed-bed reactor. Furthermore, the influence of CO on the homogeneous oxidation of HCN, possibly a product of the char-N oxidation, was evaluated using a detailed chemical kinetic model. The results indicate that CO influences the NO yield from char combustion through two paths at 850 °C: (1) CO accelerates NO reduction over char and (2) CO accelerates HCN oxidation, increasing the possibility of NO reduction over char. Both effects were more pronounced at 850 °C than at 1050−1150 °C. The present work indicates that the effect of CO 2 on NO formation in oxy-fuel combustion in fluidized beds can partly be attributed to heterogeneous reactions, whereas for high-temperature pulverized fuel combustion, CO 2 mainly affects the volatile chemistry.
Arginine-Glycine-Aspartate (RGD) tripeptide can promote cell adhesion when present in the amino acid of proteins such as fibronectin. In order to demonstrate the bioactivity of an RGD-containing silk protein, a gene encoding the RGD motif-containing peptide GSGAGGRGDGGYGSGSS (–RGD–) derived from nonmulberry silk was designed and cloned, then multimerised and inserted into a commercial pGEX expression vector for recombinant expression of (–RGD–)n peptides. Herein, we focus on two glutathione-S-transferase (GST)-tagged fusion proteins, GST–(–RGD–)4 and GST–(–RGD–)8, which were expressed in Escherichia coli BL21, purified by GST affinity chromatography, and analyzed with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MS). Target peptides (–RGD–)4 and (–RGD–)8 (6.03 and 11.5 kDa) were cleaved from the GST-tag by thrombin digestion, as verified with MS and SDS-PAGE. Isoelectric point analysis confirmed that target peptides were expressed and released in accordance with the original design. Target peptides self-assembled into a mainly α-helical structure, as determined by circular dichroism spectroscopy. Furthermore, (–RGD–)4 and (–RGD–)8 modified mulberry silk fibroin films were more effective for rapid cell adhesion, spreading and proliferative activity of L929 cells than some chemically synthesized RGD peptides modified and mulberry silk lacking the RGD motif.
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