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[en] Authors report on a carbon nanotube (CNT)-based lateral flow immunoassay (LFI) for ultrasensitive detection of proteins. Shortened multiwalled CNTs were used as a colored (black) tag. The detection antibody was covalently immobilized on the CNT surface via diimide-activated conjugation between the carboxyl groups on the CNT surface and amino groups of antibodies. The assay involved the capture of target protein in a sandwich-type format between an immobilized capture antibody on the test zone of LFI and a CNT-labelled detection antibody. CNTs were thus captured on the test zone of the LFI and gave a black colored line to enable visual detection of protein. Quantitative results were obtained by reading the test line intensities with a portable strip reader. Rabbit IgG was used as a model target to demonstrate the proof-of-concept. Combining the advantages of lateral flow assay with the unique physical properties of CNT (color, high aspect-to-size ratio and ease of surface modification), the optimized LFI can detect of 1.3 pg mL−1 of rabbit IgG (S/N = 3). This is three orders lower than that of gold nanoparticle-based LFI. Rabbit IgG spiked into human plasma samples was successfully detected with this LFI. Conceivably, this method can be extended to various other proteins for which adequate antibodies do exist. .
[en] Okadaic acid (OKA), a marine toxin produced by dinoflagellates, is responsible for most human diarrhetic shellfish poisoning-associated health disorders. A competitive displacement assay for OKA is described here. An OKA-binding aptamer was truncated with two sequences, one labeled with 6-carboxyfluorescein (FAM), and one with a quencher. On addition of OKA, it will bind to the aptamer and green fluorescence pops up because label and quencher become spatially separated. One of the truncated aptamers exhibis an excellent binding capability (Kd 2.77 nM) for OKA compared to its full-length aptamer (526 nM). The selectivity of the assay was proven by the successful fluorometric determination of OKA in the presence of common diarrhoetic toxins and in shellfish extracts. The detection limit is as low as 39 pg·mL−1. .
[en] Are define nanomaterials those materials which have at least one dimension in the range between 1 and 100 nm. By the term nanotechnology refers, instead, to the study of phenomena and manipulation of materials at the atomic and molecular level. The materials brought to the nanometric dimensions take particular chemical-physical properties different from the corresponding conventional macro materials. Speaking about the structure of nanoscale, you can check some basic properties materials (eg. Melting temperature, magnetic and electrical properties) without changing its chemical composition. In this perspective are crucial knowledge and control of production processes in order to design and get the nanomaterial more suitable for a specific application. For this purpose, it describes a series of processes of production of nanomaterials with application examples.
[it]Si definiscono nanomateriali quei materiali che hanno almeno una dimensione nell’intervallo compreso tra 1 e 100 nm. Con il termine nanotecnologia si fa riferimento, invece, allo studio dei fenomeni e della manipolazione dei materiali a livello atomico e molecolare. I materiali portati alle dimensioni nanometriche assumono particolari proprietà chimico-fisiche differenti dai corrispondenti macromateriali convenzionali. Intervenendo sulla struttura dei materiali a scala nanometrica è possibile controllarne alcune fondamentali proprietà (ad es. temperatura di fusione, proprietà magnetiche ed elettriche) senza variarne la composizione chimica. In tale ottica sono di fondamentale importanza la conoscenza e il controllo dei processi produttivi al fine di progettare e ottenere il nanomateriale più adatto a una specifica applicazione. A tale scopo vengono descritti una serie di processi di produzione dei nanomateriali con esempi applicativi
[en] The authors describe a gold nanocage-based lateral flow strip biosensor (LFSB) for low-cost and sensitive detection of IgG. This protein was used as a model analyte to demonstrate the proof-of-concept. The method combines the unique optical properties of gold nanocages (GNCs) with highly efficient chromatographic separation. A sandwich-type of immunoreactions occurs on the GNC-based LFSB which has the attractive features of avoiding multiple incubation, separation, and washing steps. The captured GNCs on the purple test zone and control zone of the biosensor are producing characteristic purple bands, and this enables IgG even to be visually detected. Quantitatation was accomplished by reading the intensities of the bands with a portable strip reader. The LFSB fabrication and assay parameters were optimized. The biosensor displays a linear response in the 0.5 to 50 ng⋅mL−1 IgG concentration range, and it has a 15 min assay time. The detection limit is 0.1 ng⋅mL−1 of IgG, which is 2.5 times lower than that when using a gold nanoparticle-based LFSB. In our perception, this assay has a wide potential for the detection of other proteins and species for which respective antibodies are available. < Image>.
[en] An electrochemiluminescence (ECL) based assay is described for the determination of the endocrine disruptor bisphenol A (BPA). The method is based on the use of carboxylated graphitic carbon nitride (C-g-C3N4) carrying an immobilized aptamer against BPA. In the presence of BPA, the ECL signal decreases due to ECL energy transfer from excited-state C-g-C3N4 to the BPA oxidation product. Under the optimal conditions, ECL intensity increases linearly in the 0.1 pM to 1 nM BPA concentration range. The detection limit is as low as 30 fM. The assay has excellent sensitivity, outstanding stability and high selectivity. It was applied to the determination of BPA in spiked water samples. .
[en] A novel chemiluminescence resonance energy transfer (CRET) system was developed and combined with a structure-switching aptamer for the highly sensitive detection of platinum. Platinum was chosen as a model analyte to demonstrate the generality of the new CRET system. This aptameric platform consisted of a streptavidin labeled aptamer against platinum and a streptavidin-coated magnetic bead for the selective separation of platinum-bound aptamer. The platinum–aptamer probe contained several guanine (G) bases bound to the 3,4,5-trimethoxyphenyl-glyoxal (TMPG) donor group at the 5′ end, a fluorescent acceptor (6-carboxy-2′,4,7,7′-tetrachlorofluorescein, TET) at the 3′ end, and a streptavidin aptamer sequence in which several base pairs were replaced by the G-G mismatch to induce the platinum-oligonucleotide coordination. The chemiluminescence (CL) generated by TMPG/G bases is transferred to the acceptor (TET). In the presence of platinum, the platinum–aptamer probe was folded such that the G bases at the 5′ end and TET at the 3′ were in close proximity. The complex was separated using streptavidin-coated magnetic beads by the addition of TMPG to form the TMPG/G bases complex. The ultraweak CL from the TMPG/G bases was strongly enhanced by TET. This novel CRET-based method can be easily performed with high limit of detection (50 ng·mL−1) and selectivity over other metal ions. This technique provides a novel method for simple, fast, and convenient point-of-care diagnostics for monitoring proteins and metal ions. .
[en] A highly sensitive fluorometric method is described for the determination of mercury(II) ions. It is based on (a) the use of a DNA probe containing thymine-thymine mismatches that are employed as Hg(II) recognition elements, (b) subsequent toehold binding, and (c) endocuclease-assisted signal amplification. Target recycling is triggered by exonuclease III. This produces a large amount of ssDNA (defined as primer). Then, the generated primer-initiated strand displacement reaction with the help of polymerase and nicking endonuclease releases the free fluorophore-labelled probe. Under excitation at 532 nm, the fluorescent probe displays emission with a peak at 582 nm. The sensitivity of this method is improved by introduction of nicking endonuclease. The working range of the assay extends from 20 pM to 10 nM, and the detection limit is as low as 6 pM of Hg(II). .
[en] Preparation of copper ferrite polyaniline nanocomposite (CF-PANI) and its use for photocatalytic degradation of Rhodamine B dye from aqueous solution has been studied. Copper ferrite (CuFe2O4), a magnetic ferrite (spinel) of nano dimension was synthesized by co-precipitation method. Polyaniline nanocomposite of copper ferrite was prepared by in situ polymerization and SEM technique was used for characterization. Adsorption and photo-degradation capacity of prepared nanocomposite was evaluated under different conditions. (author)
[en] The authors describe an isothermal and ultrasensitive colorimetric DNA assay that consists of two amplification stages using enzymes and a catalytic hairpin assembly (CHA). The first step consists in the selective amplification of DNA using Klenow fragment and nicking enzyme. The second step consists in the amplification of the optical signal by using a catalytic hairpin assembly. After two amplification steps, the DNA reaction induces the aggregation of the red gold nanoparticles to give a blue color shift. The degree of aggregation can be quantified by measurement of the ratio of the UV-vis absorbances of the solutions at 620 and 524 nm which are the wavelengths of the aggregated gold nanoparticles and bare gold nanoparticles. The detection limit is as low as 3.1 fM. Due to the use of a specific enzyme, only the desired DNAs will be detected. The method can be applied to the determination of DNA of various lengths. Despite the presence of large amounts of wildtype DNA, it can readily detect a target DNA. Conceivably, the technique has a large potential because of its high sensitivity and selectivity. .