查看更多>>摘要：Metal nanoclusters (NCs) as promising nanomaterials for sensing applications have attracted significant attention because of their unique photoluminescence properties. However, the quantum yields of metal NCs are still relatively low when compared to conventional quantum dots and organic dyes, posing a major obstacle to their assay application. It is challenging but important to pursue a way to improve the luminescence of metal NCs. In this work, we developed a novel strategy to enhance the luminescence of silver nanoclusters (Ag NCs) based on the binding with 6-aza-2-thiothymine (ATT) via Au3+ bridging. We studied the possible mechanism of this binding-induced luminescence enhancement and attributed it to the ligands rigidifying. Since 2-thiouracil (2TU), a common anticancer, antithyroid, and antiviral agent, featured a similar molecular structure of ATT, this luminescence enhancement strategy can be designed to sensitive and selective turn-on detect 2-TU. As far as we know, this is the first report for the fluorescent turn-on detect 2-TU. Benefiting from the good performance of this method and the advantages of fluorescence assay, intracellular imaging of 2-TU, which has yet to be achieved based on currently developed analytical methods for 2-TU, was carried out via our approach. Moreover, to further expand the sensing application of the developed luminescence enhancement method, we constructed a universal detection platform. Taking hyaluronidase as a target, the feasibility of the detection platform was confirmed. The discoveries in this study offer a simple route to improve the optical properties of NCs and design their sensing applications.
查看更多>>摘要：Nucleocapsid protein (N protein) is the most abundant protein in SARS-CoV2 and is highly conserved, and there are no homologous proteins in the human body, making it an ideal biomarker for the early diagnosis of SARS-CoV2. However, early detection of clinical specimens for SARS-CoV2 remains a challenge due to false-negative results with viral RNA and host antibodies based testing. In this manuscript, a microfluidic chip with femtoliter-sized wells was fabricated for the sensitive digital detection of N protein. Briefly, beta-galactosidase (beta-Gal)-linked antibody/N protein/aptamer immunocomplexes were formed on magnetic beads (MBs). Afterwards, the MBs and beta-Gal substrate fluorescein-di-beta-D-galactopyranoside (FDG) were injected into the chip together. Each well of the chip would only hold one MB as confined by the diameter of the wells. The MBs in the wells were sealed by fluorocarbon oil, which confines the fluorescent (FL) product generated from the reaction between beta-Gal and FDG in the individual femtoliter-sized well and creates a locally high concentration of the FL product. The FL images of the wells were acquired using a conventional inverted FL microscope. The number of FL wells with MBs (FL wells number) and the number of wells with MBs (MBs wells number) were counted, respectively. The percentage of FL wells was calculated by dividing (FL wells number) by (MBs wells number). The higher the percentage of FL wells, the higher the N protein concentration. The detection limit of this digital method for N protein was 33.28 pg/mL, which was 300 times lower than traditional double-antibody sandwich based enzyme-linked immunosorbent assay (ELISA).
查看更多>>摘要：Real time controllable assembling/aptasensing approach via plasmonic graphene oxide (GO) nanocomposites has been firstly proven to simultaneously give tuning of micro-nano structure of plasmonic GO and ultrasensitive detection of MC-LR toxin. In order to fabricate the assembly, a high-quality hollow triangular nanoplate AgClAu: p-GO (HTNP AgClAu:p-GO) can act as a template; furthermore, we combine DNA-hybridization with biotinstrepavidin binding protocol for tuning the HTNP AgClAu:p-GO assemblies from networks to laminar structure, and simultaneously loading Raman reporters into the assemblies. The dynamic assembling process can be utilized as a real time SERS aptasensor for detecting MC-LR due to ratiometric introduction of MC-LR toxin inhibiting formation of plasmonic p-GO assembly via toxin/aptamer bioconjugation and causing reverse alteration of SERS signal for giving ultrasensitive SERS detection of MC-LR. A detection limit of 6.3pM with a wide linear range from 10pM to 5 nM can be achieved. When the aptasensor has been applied in real samples, the real time assembling/aptasensing approach shows recoveries from 98% to 103% with relative standard deviation (RSD) lower than 3%, expecting that one-step nanofabrication and sensing strategy can be extended to in-field test of environmental contaminants.
查看更多>>摘要：In this study, a novel type of covalent organic framework (COF) material rich in boronic acid sites was prepared through post-synthesis modification (TbBD@PEI@Au@4-MPBA). The surface of COF material had abundant carboxylic acid groups, which could bind a large amount of polyethyleneimine (PEI) through electrostatic interaction. At the same time, the amino groups on the PEI can be grafted with Au nanoparticles (Au NPs) in situ, and then 4-mercaptophenylboronic acid (4-MPBA) was modified by the reaction of Au and sulfhydryl groups. The massive grafting of boronic acid groups made the material's enrichment effect on glycopeptides expected. The results of experiments indicated that the composite material has high sensitivity (5 amol mu L-1) and selectivity (1:1000). In addition, the material has outstanding stability and reusability, with a load capacity of about 100 mg g-1 and a recovery of 99.3 +/- 2.2%. What's more, after enriched by TbBD@PEI@Au@4-MPBA, 56 endogenous glycopeptides from fresh human saliva were detected by MALDI-TOF MS, 56 unique glycopeptides corresponding to 31 glycoproteins from human saliva and 513 unique glycopeptides corresponding to 208 glycoproteins from serum of throat cancer patient were detected by nano-LC-MS/MS, respectively, which was expected to be applied to glycoproteomics research.
Kasperkiewicz, AlexanderLendor, SofiaPawliszyn, Janusz
6页
查看更多>>摘要：Relative matrix effects between an ambient mass spectrometric technique known as coated blade spray (CBS) and liquid chromatographic separation approach when applied to multiresidue pesticide analysis in strawberry samples are explored. Acceptable slope relative standard deviations (RSD <15 %) were observed for the 9 compounds under study for both CBS-MS/MS (2.2-12.6 %) and LC-MS/MS (2.8-12.9 %) approaches. The findings signify both the elimination of relative matrix effects with the sample preparation and matrix match calibration with internal standard correction methods employed along with no matrix effect compromise made when using the direct-to-MS approach. Similarly, slopes of pesticides spiked from commercially available formulations (containing one or two pesticides) were found to not differ significantly from slopes generated with multiresidue pesticide standards (containing 24 additional pesticides besides the target 9 analytes) with either technique, highlighting the resistance of the employed methods to the excipients present in pesticide formulations in large amounts.
查看更多>>摘要：Lipidomics has great potential for the discovery of biomarkers, elucidation of metabolic processes and identifying dysregulations in complex biological systems. Concerning biofluids like plasma or cerebrospinal fluid, several studies for the comparison of lipid extraction solvents have already been conducted. With respect to tissues, which can differ significantly in terms of dry matter content and composition, only few studies are available. The proper selection of an extraction method that covers the complexity and individuality of different tissues is challenging. The goal of this work was to provide a systematic overview on the potential of different extraction methods for a broad applicability. This study covers six different extraction procedures and four different reconstitution solvents applied to ten different porcine tissues. To get an overview of the individual lipid profiles, a workflow was developed for a fast and reliable tentative lipid annotation. Therefore, several machine learning tools were utilized, like the prediction of collision cross sections to support the tentative lipid identification in case of untargeted lipidomics. In terms of data evaluation, unsupervised (e.g. principal component analysis) and supervised (e.g. partial least square - discriminant analysis) methods were applied to visualize and subsequently interpret all generated information. Furthermore, the influence of the tissue composition on the extraction performance was investigated. It could be shown that the ten porcine tissues can be distinguished based on their lipid profile with the applied workflow and that the methyl-tert-butyl ether (MTBE) based extraction method (two-phase) showed the best overall performance for the 16 examined lipid species. With this method the highest number of features (428 in lung tissue) could be annotated. Upcoming one-phase extractions also showed a high potential concerning total number of extracted lipids. Methanol/MTBE/chloroform (MMC) extracted slightly less lipids (393 in lung and liver) than MTBE but turned out to be the best one-phase extraction method. Additionally, the numbers of extracted lipids obtained by isopropanol/water 90:10 (IPA90) (399 in stomach) and by isopropanol/methanol/chloroform (IMC) (395 in stomach) were similar to those of the modified Folch method (402 in stomach). One-phase extractions can therefore clearly be seen as preferable when a high throughput is needed.
查看更多>>摘要：A fluorescent aptasensor based on sensitized terbium(III) luminescence was constructed to detect melamine in milk. Tb3+ as the fluorescence probe can be sensitized by a guanine-rich single-stranded DNA sequence, so the complementary sequence of the polythymidine aptamer (cDNA) was modified with six consecutive guanine bases (G6). In the absence of melamine, melamine aptamer combined with cDNA to form a double helix structure, and G6 hybridized with the extended cytosine bases in the aptamer, resulting in low fluorescence intensity of Tb3+. In the presence of melamine, cDNA was released due to the specific recognition of melamine to the aptamer, resulting in stronger sensitized fluorescence intensity of Tb3+. Under the optimum conditions, the linear concentration of melamine in the milk ranged from 1.0 mu g/mL to 10.0 mu g/mL. This aptasensor can be used for the accurate and rapid detection of melamine in milk with a detection limit of 0.02 mu g/mL, and has the advantages of high sensitivity, high efficiency, simple operation and low cost.
查看更多>>摘要：A real-time quartz crystal microbalance (QCM) cytosensor was first developed for dynamical and noninvasive monitoring of cell viscoelasticity for evaluation of apoptosis degree. In this work, human breast cancer cells MCF-7 and MDA-MB-231 were employed as cell model and respectively captured on the surface of QCM electrode modified with mercaptosuccinic acid and poly-L-lysine. Cell viscoelasticity was measured dynamically by real-time monitoring energy dissipation with QCM, and the dynamic diagram of the energy dissipation of MDA-MB-231 cells treated with curcumin was first obtained. The results displayed that the changes of energy dissipation in MDA-MB-231 cells and MCF-7 cells were 8.81 x 10(-6) and 5.29 x 10(-6), particularly due to the difference in cell viscoelasticity. Furthermore, curcumin was used to induce cell apoptosis and suppress energy dissipation of MDA-MB-231 cells. Combining apoptosis assay with QCM measurement, the results revealed good linear relationship between cell viscoelasticity inhibition and apoptosis rate with correlation coefficient R = 0.9908. The QCM cytosensor could rapidly, accurately, dynamically, and noninvasively monitor the changes of cell viscoelasticity for evaluation of apoptosis degree in MDA-MB-231 cells. The study established a new model for cell apoptosis assessment, facilitating understanding of the mechanisms of cell apoptosis on the aspect of mechanical properties.
查看更多>>摘要：Bacillus thuringiensis (Bt) is used as a bioinsecticide since it effectively kills insect larvae. Bt is also genetically similar to Bacillus cereus (Bc), a well recognized foodborne human pathogen; they are both members of the Bacillus cereus group (BC group). Although approved Bt bioinsecticide products have been confirmed to be nonpathogenic to humans, close monitoring of Bt during dissemination is important for cost considerations and to limit impact on biodiversity towards nontarget organisms. As such, developing rapid, sensitive, and specific tools for quantitative detection of Bt spores during and following spray operations is highly desirable. The goals of this study were to investigate commercially available detection reagents for sensitivity and selectivity in detecting Bt spores, and then functionalize a surface of (001) GaAs used in photonic biosensing. To achieve these goals, we (1) screened commercial antibodies for their capacity to bind recombinant proteins from Bt spores, (2) screened antibodies and aptamers for their sensitivity and selectivity against Bt spores, and (3) tested the efficiency of selected antibodies and aptamers in capturing Bt spores on the surface of functionalized GaAs biochips. Seven genes encoding Bt spore proteins were cloned and expressed in Escherichia coli. The binding of each purified spore antigen was tested by commercially available polyclonal and monoclonal antibodies claimed to exclusively target spores. Of the seven targets, Bacillus collagen-like protein A, was the most abundant protein on Bt spores and demonstrated the strongest binding affinity to all test antibodies. The commercial antibodies (Abs) were also tested for specificity to BC Group versus non-BC Group spores. Three of six commercial antibodies showed selectivity to Bt spores, with recombinant Abs providing the most robust lower range of detection (102 to 6 x 103 spores/mL). The sensitivity and selectivity of three published DNA aptamer sequences demonstrated a wide range of detection sensitivity for Bt spores. Two of the three test aptamers also showed reasonable selectivity towards Bt spores while the third demonstrated reactivity to non-BC Group B. megaterium and B. subtilis. Of the reagents tested, a thiolated aptamer and llama recombinant Ab showed highest Bt spore capture efficiency as measured by spore coverage of the GaAs surface. These results confirm that the selected aptamer and llama rAb can be considered strong candidates for the development of GaAs-based biosensing devices.
NSTL
Elsevier