Amplification of an Electrochemiluminescence-Emissive Aptamer into DNA Nanotags for Sensitive Fecal Calprotectin Determination.

The precision additive manufacturing and tessellated multitasking out of the structural DNA nanotechnology enable a configurable expression of densified electrochemiluminescent (ECL) complexes, which would streamline the bioconjugation while multiplying signals. Herein, a completely DNA-scaffold ECL "polyploid" was replicated out via the living course of rolling circle amplification. The amplicon carried the aptameric sequences of ZnPPIX/TSPP porphyrin as photoreactive centers that rallied at periodical intervals of the persistent extension into a close-packed nanoflower, ZnPDFI/II. Both microscopies and electrophoresis proved the robust nesting of guests at their deployed gene loci, while multispectral comparisons among cofactor substituents pinpointed the pivotal roles of singlet seclusion and Zn2+-chelation for the sake of intensive ECL irradiation. The adversity-resilient hydrogel texture made lipoidal filmogens as porphyrinic ECL prerequisites to be of no need at all, thus not only simplifying assay flows but also inspiring an in situ labeling plan. Upon bioprocessing optimization, an enriched probe ZnPDFIII was further derived that interpolated the binding motif related to calprotectin as validated by molecular docking and affinity titration. With it being a strongly indicative marker of inflammatory bowel disease (IBD), a competitive ECL aptasensing strategy was contrived, managing a signal-on and sensitive detection in mild conditions with a subnanogram-per-milliliter limit of detection by 2 orders of magnitude lower than the standard method as well as a comparable accuracy in clinical stool sample testing. Distinct from those conventional chemophysical rebuilding routes, this de novo biosynthetic fusion demonstrated a promising alternative toward ECL-source bioengineering, which may intrigue vibrant explorations of other ECL-shedding fabrics and, accordingly, a new bioanalytic mode downstream.

In situ formed copper nanoparticles templated by TdT-mediated DNA for enhanced SPR sensor-based DNA assay.

For the efficient surface plasmon resonance (SPR)-based DNA assay researching, signal amplification tactics were absolutely necessary. In this work, a sensitive SPR-DNA sensor was developed by employing in situ synthesis of copper nanoparticles (CuNPs) templated by poly-T sequences DNA from terminal deoxynucleotidyl transferase (TdT)-mediated extension, and synergistically with nano-effect deposition as the mass relay. The objective of this strategy was manifold: firstly, tDNA hybridized with the optimal designed probes to active the TdT-mediated DNA extension onto the surface of SPR chip, resulted a long poly-T sequences ssDNA chain in dsDNA terminal onto surface of gold chip and characterized by SPR signal amplitudes. Secondly, copper ion (Cu2+) adsorbed into the skeleton of poly-T sequences DNA, with the aid of ascorbic acid (VC) to achieve the Cu2+ reduction, copper nanostructures (CuNPs) was synchronously generated onto the single nucleotide chain anchoring in dsDNA derivatives and the formation was featured by transmission electron micrographs (TEM) and electrochemistry. Lastly, dsDNA-complexed CuNPs (CuNPs@dsDNA) triggered the final signal amplification via real-time conversion of the additive catechol violet (CV) into oligomer or chelation precipitation by CuNPs-tagged reporters. With the proposed setups, a precise and replicable DNA sensing platform for specific target oligo was obtained with a detection limit down to 3.21 femtomolar, demonstrating a beneficial overlapping exploitation of nanomaterials and biochemical reaction as unique SPR infrastructure. Such triple-amplification strategic setups, the possibility of various methods abutment and biocompatibility weight reactor was amassed and adapted to more biological detection field.

Polymerization amplified SPR-DNA assay on noncovalently functionalized graphene.

A highly efficient surface plasmon resonance (SPR)-based DNA assay was developed, by employing noncovalently functionalized graphene nanosheets as a substrate, and enzymatic catalysis-induced polymerization as mass relay. The objective of this strategy was manifold: first of all, to sensitize the overall SPR output by in situ optimized electrogeneration of graphene thin-film, which was characterized by atomic force microscopic topography; secondly, to regulate the self-assembly and orientation of biotinylated capture probes on nickel-chelated nitrilotriacetic acid (NTA) scaffolds, that anchored onto graphene-supported pyrenyl derivatives; and lastly, to synergize the signal amplification via real-time conversion of the additive aniline into polyaniline precipitation by horseradish peroxidase-tagged reporters. With this setup, a precise and replicable DNA sensing platform for specific targets was achieved with a detection limit down to femtomolar, thus demonstrating a beneficial exploration and exploitation of two-dimensional nanomaterials as unique SPR infrastructure. The possibility of such ″bottom-up″ architecture mounted with ″top-down″ weight reactor would be most likely extensible and adaptable to protein determinations.

Magnetic zirconium hexacyanoferrate(II) nanoparticle as tracing tag for electrochemical DNA assay.

Novel multifunctional magnetic zirconium hexacyanoferrate nanoparticles (ZrHCF MNPs) were prepared, which consisted of magnetic beads (MBs) inner core and zirconium hexacyanoferrate(II) (ZrHCF) outer shell. As an artificial peroxidase, the ZrHCF MNPs exhibited remarkable electrocatalytic properties in the reduction of H2O2 at 0.2 V vs saturated calomel electrode (SCE). On the basis of the bonding interaction between Zr (IV) of the shell ZrHCF framework and phosphonate groups, the 5'-phosphorylated ssDNA probes with a consecutive stretch of guanines as a spacer could be incorporated in ZrHCF MNPs easily. Thus, DNA-grafted ZrHCF MNPs could be simply obtained by magnetic separation. The prepared nanoelectrocatalyst was further used as signal nanoprobe for the ultrasensitive electrochemical DNA assay. Under optimal conditions, the proposed biosensor presents high sensitivity for detecting target DNA with a linear range from 1.0 fM to 1.0 nM and a low detection limit of 0.43 fM. Moreover, it exhibits good performance with excellent selectivity, high stability, and acceptable fabrication reproducibility.

Mass effect of redox reactions: A novel mode for surface plasmon resonance-based bioanalysis.

The pursuit of more specific and sensitive response is a perpetual goal for modern bioassays. This work proposed a novel label-free strategy about redox-related mass effect based on the surface plasmon resonance (SPR) technique for ultrasensitive determination of DNA. The protocol starts with the modification of SPR gilded disk with the capture DNA (cDNA). After the conjugation of immobilized cDNA with the target DNA (tDNA), the hybridization chain reaction was triggered by the introduction of mutual partial complementary primers to elongate the terminal into a nanoscale duplex. As it is reported that porphyrin could intercalate into the grooves of the double-stranded DNA (dsDNA) scaffold, multiple positive-charged Fe(III)meso-tetra(N-methyl-4-pyridyl) porphine (FeTMPyP) with symmetric structure were uptaken for in situ formation of porphyrin-dsDNA complex. Given FeTMPyP a highly efficient catalysis for the peroxide reduction, its presence as a biomimetic cofactor was validated via circular dichroism and UV-vis spectroscopy, demonstrating a tight binding as well as high catalytic activity and stability. Using 4-chloro-1-naphthol as a proton donor, the catalytic reduction of H2O2 would oxidize it into insoluble benzo-4-chloro-hexadienone, which simultaneously deposited on the heterogeneous interface, leading to a significant amplification in both SPR response and topological height profile. The signal increment was proportional to the concentration of tDNA, thus an ultrasensitive SPR-based DNA assay was developed with a linear range over four orders of magnitudes and a sub-femtomolar detection limit of 0.73 fM. The developed methodology exemplifies a different way of thinking about mass-sensing modes, extending conventional SPR-based DNA analysis to relevant biomedical applications.

Unusual Fe(CN)6³â»/4⁻ capture induced by synergic effect of electropolymeric cationic surfactant and graphene: characterization and biosensing application.

Herein, a special microheterogeneous system for Fe(CN)6(3-/4-) capture was constructed based on graphene (GN) and the electropolymeric cationic surfactant, an amphiphilic pyrrole derivative, (11-pyrrolyl-1-yl-undecyl) triethylammonium tetrafluoroborate (A2). The morphology of the system was characterized by scanning electron microscope. The redox properties of the entrapped Fe(CN)6(3-/4-) were investigated by cyclic voltammetry and UV-visible spectrometry. The entrapped Fe(CN)6(3-/4-) exhibited highly electroactive with stable and symmetrical cyclic voltammetric signal. A dramatic negative shift in the half wave potential can be obtained due to the unusual Fe(CN)6(3-/4-) partitioning in in this microheterogeneous system based on poly(A2+GN). Finally, the entrapped Fe(CN)6(3-/4-) was applied in the construction of the enhanced biosensors to hydrogen peroxide and sulfide.

Bioinspired polydopamine as the scaffold for the active AuNPs anchoring and the chemical simultaneously reduced graphene oxide: characterization and the enhanced biosensing application.

We report here an efficient approach to enhance the performance of biosensing platform based on graphene or graphene derivate. Initially, graphene oxides (GO) nanosheets were reduced and surface functionalized by one-step oxidative polymerization of dopamine in basic solution at environment friendly condition to obtain the polydopamine (Pdop) modified reduced graphene oxides (PDRGO). The bioinspired surface was further used as a support to anchor active gold nanoparticles (AuNPs). The morphology and structure of the as-prepared AuNPs/PDRGO nanocomposite were investigated by field-emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), Fourier transform-infrared spectroscopy (FT-IR). Electrochemical studies demonstrate that the as-prepared AuNPs/PDRGO hybrid materials possess excellent electrochemical properties and electrocatalytic activity toward the oxidation of NADH at low potential (0.1 V vs. SCE) with the fast response (15s) and the broad linear range (5.0 × 10(-8)-4.2 × 10(-5)M). Thus, this AuNPs/PDRGO nanocomposite can be further used to fabricate a sensitive alcohol biosensor using alcohol dehydrogenase (ADH), by simply incorporating the specific enzyme within the composite matrix with the aid of chitosan (Chit).