On‐the‐Spot Immobilization of Quantum Dots,Graphene Oxide,and Proteins via Hydrophobins

Class I hydrophobin Vmh2, a peculiar surface active and versatile fungal protein, is known to self‐assemble into chemically stable amphiphilic films, to be able to change wettability of surfaces, and to strongly adsorb other proteins. Herein, a fast, highly homogeneous and efficient glass functionalization by spontaneous self‐assembling of Vmh2 at liquid–solid interfaces is achieved (in 2 min). The Vmh2‐coated glass slides are proven to immobilize not only proteins but also nanomaterials such as graphene oxide (GO) and quantum dots (QDs). As models, bovine serum albumin labeled with Alexa 555 fluorophore, anti‐immunoglobulin G antibodies, and cadmium telluride QDs are patterned in a microarray fashion in order to demonstrate functionality, reproducibility, and versatility of the proposed substrate. Additionally, a GO layer is effectively and homogeneously self‐assembled onto the studied functionalized surface. This approach offers a quick and simple alternative to immobilize nanomaterials and proteins, which is appealing for new bioanalytical and nanobioenabled applications.     

Tunable electrochemistry of gold-silver alloy nanoshells

Nano Research - The widespread and increasing interest in enhancing biosensing technologies by increasing their sensitivities and lowering their costs has led to the exploration and application of...     

Double-codified gold nanolabels for enhanced immunoanalysis

A novel double-codified nanolabel (DC-AuNP) based on gold nanoparticle (AuNP) modified with anti-human IgG peroxidase (HRP)-conjugated antibody is reported. It represents a simple assay that allows enhanced spectrophotometric and electrochemical detection of antigen human IgG as a model protein. The method takes advantage of two properties of the DC-AuNP label: first, the HRP label activity toward the OPD chromogen that can be related to the analyte concentration and measured spectrophotometrically; second, the intrinsic electrochemical properties of the gold nanoparticle labels that being proportional to the protein concentration can be directly quantified by stripping voltammetry. Beside these two main direct determinations of human IgG, a secondary indirect detection was also applicable to this system, exploiting the high molar absorptivity of gold colloids, by which, the color intensity of their solution was proportional to the concentration of the antigen used in the assay. Paramagnetic beads were used as supporting material to immobilize the sandwich-type immunocomplexes resulting in incubation and washing times shorter than those typically needed in classical ELISA tests by means of a rapid magnetic separation of the unbound components. A built-in magnet graphite-epoxy-composite electrode allowed a sensibly enhanced adsorption and electrochemical quantification of the specifically captured AuNPs. The used DC-AuNP label showed an excellent specificity/selectivity, as a matter of fact using a different antigen (goat IgG) a minimal nonspecific electrochemical or spectrophotometric signal was measured. The detection limits for this novel double-codified nanoparticle-based assay were 52 and 260 pg of human IgG/mL for the spectrophotometric (HRP-based) and electrochemical (AuNP-based) detections, respectively, much lower than those typically achieved by ELISA tests. The developed label and method is versatile, offers enhanced performances, and can be easily extended to other protein detection schemes as well as in DNA analysis.     

Glucose biosensor based on carbon nanotube epoxy composites

A novel glucose biosensor based on a rigid and renewable carbon nanotube (CNT) based biocomposite is reported. The biosensor was based on the immobilization of glucose oxidase (GOx) within the CNT epoxy-composite matrix prepared by dispersion of multi-wall CNT inside the epoxy resin. The use of CNT, as the conductive part of the composite, ensures better incorporation of enzyme into the epoxy matrix and faster electron transfer rates between the enzyme and the transducer. Experimental results show that the CNT epoxy composite biosensor (GOx-CNTEC) offers an excellent sensitivity, reliable calibration profile, and stable electrochemical properties together with significantly lower detection potential (+0.55 V) than GOx-graphite epoxy composites (+0.90 V; difference deltaE = 0.35 V). The results obtained favorably compare to those of a glucose biosensor based on a graphite epoxy composite (GOx-GEC).     

Toward integrated detection and graphene-based removal of contaminants in a lab-on-a-chip platform

A novel miniaturized microfluidic platform was developed for the simultaneous detection and removal of polybrominated diphenyl ethers (PBDEs).The platform consists of a polydimethylsiloxane (PDMS) microfluidic chip for an immunoreaction step,a PDMS chip with an integrated screen-printed electrode (SPCE) for detection,and a PDMS-reduced graphene oxide (rGO) chip for physical adsorption and subsequent removal of PBDE residues.The detection was based on competitive immunoassay-linked binding between PBDE and PBDE modified with horseradish peroxidase (HRP-PBDE) followed by the monitoring of enzymatic oxidation of o-aminophenol (o-AP) using square wave anodic stripping voltammetry (SW-ASV).PBDE was detected with good sensitivity and a limit of detection similar to that obtained with a commercial colorimetric test (0.018 ppb),but with the advantage of using lower reagent volumes and a reduced analysis time.The use of microfluidic chips also provides improved linearity and a better reproducibility in comparison to those obtained with batch-based measurements using screen-printed electrodes.In order to design a detection system suitable for toxic compounds such as PBDEs,a reduced graphene oxide-PDMS composite was developed and optimized to obtain increased adsorption (based on both the hydrophobicity and π-π stacking between rGO and PBDE molecules) compared to those of non-modified PDMS.To the best of our knowledge,this is the first demonstration of electrochemical detection of flame retardants and a novel application of the rGO-PDMS composite in a biosensing system.This system can be easily applied to detect any analyte using the appropriate immunoassay and it supports operation in complex matrices such as seawater.     

In Situ Production of Biofunctionalized Few‐Layer Defect‐Free Microsheets of Graphene

Biological interfacing of graphene has become crucial to improve its biocompatibility, dispersability, and selectivity. However, biofunctionalization of graphene without yielding defects in its sp²‐carbon lattice is a major challenge. Here, a process is set out for biofunctionalized defect‐free graphene synthesis through the liquid phase ultrasonic exfoliation of raw graphitic material assisted by the self‐assembling fungal hydrophobin Vmh2. This protein (extracted from the edible fungus Pleurotus ostreatus) is endowed with peculiar physicochemical properties, exceptional stability, and versatility. The unique properties of Vmh2 and, above all, its superior hydrophobicity, and stability allow to obtain a highly concentrated (≈440–510 μg mL^?1) and stable exfoliated material (ζ‐potential, +40/+70 mV). In addition controlled centrifugation enables the selection of biofunctionalized few‐layer defect‐free micrographene flakes, as assessed by Raman spectroscopy, atomic force microscopy, scanning electron microscopy, and electrophoretic mobility. This biofunctionalized product represents a high value added material for the emerging applications of graphene in the biotechnological field such as sensing, nanomedicine, and bioelectronics technologies.