Rapid development in technology and society has generated diverse developments in many fields including biosensors in healthcare application. Here, the design of integrated biosensor comprises mass sensing (Quartz Crystal Microbalance) and electrochemistry sensing (Electrochemical Impedance Spectroscopy, EIS and Cyclic Voltammetry, CV) will be presented. The integrated sensor system is developed based on the innovative use of the top electrode of a quartz crystal microbalance (QCM) resonator as a working electrode for the electrochemistry technique. Integration of QCM with the electrochemistry technique is realized by fabricating a semicircular counter electrode near the upper electrode on the same side of the quartz crystal. CV and EIS measurement was conducted using finite element modeling, COMSOL (TM) 5.2 with the probe marker of 1 mmol L-1 of [Fe(CN)(6)](3-/4-). CV test was done to study the effect between increasing scan rate and peak current (anodic and cathodic) in observing the reversible electrochemical process. This observation is crucial in ensuring the electrochemical processes follow the Randles-Sevcik equation in characterizing the platform changes behavior. Later, EIS test was performed in order to measure the radius of the semicircle which reflects the charge transfer resistance (R-CT) of the redox marker. To show the effectiveness of this sensor, gold immobilization surface was electrochemically simulated and reported. Thus, an ultra-sensitive biosensor that capable to produce multi-analysis in the detection of biological targets in terms of electrochemical change of electrode interfaces, which is the crucial step towards the engineering of advanced bioelectronics.

We present a low-cost, accessible, and rapid fabrication process for electrochemical microfluidic sensors. This work leverages the accessibility of consumer-grade electronic craft cutters as the primary tool for patterning of sensor electrodes and microfluidic circuits, while commodity materials such as gold leaf, silver ink pen, double-sided tape, plastic transparency films, and fabric adhesives are used as its base structural materials. The device consists of three layers, the silver reference electrode layer at the top, the PET fluidic circuits in the middle and the gold sensing electrodes at the bottom. Separation of the silver reference electrode from the gold sensing electrodes reduces the possibility of cross-contamination during surface modification. A novel approach in mesoscale patterning of gold leaf electrodes can produce generic designs with dimensions as small as 250 mu m. Silver electrodes with dimensions as small as 385 mu m were drawn using a plotter and a silver ink pen, and fluid microchannels as small as 300 mu m were fabricated using a sandwich of iron-on adhesives and PET. Device layers are then fused together using an office laminator. The integrated microfluidic electrochemical platform has electrode kinetics/performance of Delta E-p = 91.3 mV, I-pa/I-pc = 0.905, characterized by cyclic voltammetry using a standard ferrocyanide redox probe, and this was compared against a commercial screen-printed gold electrode (Delta E-p = 68.9 mV, I-pa/I-pc = 0.984). To validate the performance of the integrated microfluidic electrochemical platform, a catalytic hydrogen peroxide sensor and enzyme-coupled glucose biosensors were developed as demonstrators. Hydrogen peroxide quantitation achieves a limit of detection of 0.713 mM and sensitivity of 78.37 mu A mM(-1) cm(-2), while glucose has a limit of detection of 0.111 mM and sensitivity of 12.68 mu A mM(-1) cm(-2). This rapid process allows an iterative design-build-test cycle in under 2 hours. The upfront cost to set up the system is less than USD 520, with each device costing less than USD 0.12, making this manufacturing process suitable for low-resource laboratories or classroom settings.

Current electrochemical biosensors for multiple miRNAs require tedious immobilization of various nucleic acid probes. Here, we demonstrate an innovative approach using a generic neutravidin biosensor combined with electrochemically encoded responsive nanolabels for facile and simultaneous multiplexed detection of miRNA-21 and miRNA-141. The selectivity of the biosensor arises from the intrinsic properties of the electrochemically encoded responsive nanolabels, comprising biotinylated molecular beacons (biotin-MB) and metal nanoparticles (metal-NPs). The procedure is a simple one-pot assay, where the targeted miRNA causes the opening of biotin-MB followed by capturing of the biotin-MB-metal-NPs by the neutravidin biosensor and simultaneous detection of the captured metal-NPs by stripping square-wave voltammetry (SSWV). The multiplexed detection of miRNA-21 and miRNA-141 is achieved by differentiation of the electrochemical signature (i.e., the peak current) for the different metal-NP labels. The biosensor delivers simultaneous detection of miRNAs with a linear range of 0.5-1000 pM for miRNA-21 and a limit of detection of 0.3 pM (3 sigma/sensitivity, n = 3), and a range of 50-1000 pM for miRNA-141, with a limit of detection of 10 pM. Furthermore, we demonstrate multiplexed detection of miRNA-21 and miRNA-141 in a spiked serum sample.

Alteration in expression of miRNAs has been correlated with different cancer types, tumour stage and response to treatments. In this context, a structurally responsive oligonucleotide-based electrochemical impedimetric biosensor has been developed for the simple and sensitive detection of miRNA-21. A highly specific biotinylated DNA/LNA molecular beacon (MB) probe was conjugated with gold nanoparticles (AuNPs) to create an integrated, dual function bio-label (biotin-MB-AuNPs) for both biorecognition and signal generation. In the presence of target miRNA-21, hybridisation takes place resulting in the "activation" of the biotin-MB; this event makes the biotin group, which was previously "protected" by the steric hindrance of the MB stem-loop structure, accessible. The activated biotin-MB-AuNPs/miRNA complexes become available for capture, via supramolecular interaction, onto a nentravidin-modified electrode for electrochemical transduction. The binding event results in a decrease of the charge transfer resistance at the working electrode/electrolyte interface. The biosensor responded linearly in the range 1-1000 pM of miRNA-21, with a limit of detection of 0.3 pM, good reproducibility (Relative Standard deviation (RSD) =3.3%) and high selectivity over other miRNAs (i.e. miRNA221 and miRNA-205) sequences. Detection of miRNA-21 in spiked serum samples at clinically relevant levels (low pM range) was also demonstrated, thus illustrating the potential of the biosensor for point-of-care clinical applications. The proposed biosensor design, based on the combination of a neutravidin transducing surface and the dual-function biotin-MB-AuNPs bio-label, provides a simple and robust approach for detection of short-length nucleic acid targets, such as miRNAs.

The fabrication of inflated Layer-by-Layer (LbL) microcapsules with a unique ‘bead-in-a-capsule’ morphology is presented. Currently, the fabrication of LbL microcapsules using conventional aqueous LbL techniques usually results in microcapsules with a two-phase system (LbL capsular wall with an air, liquid, solid or hydrogel core). Here, we present the fabrication of inflated LbL microcapsules with a unique three-phase system (LbL capsular wall, hydrogel microbead in an aqueous core) by using the Reverse-Phase LbL (RP-LbL) technique. The RP-LbL technique is performed in an organic solvent and allows encapsulation of water-soluble templates and molecules with high efficiency. Firstly, the RP-LbL technique is used to coat polymer layers onto agarose microbeads containing TRIS buffer for the formation of LbL capsular walls onto the microbeads and to minimize out-diffusion of encapsulated TRIS. Next, the polymer-coated agarose microbeads are transferred from an organic to an aqueous solvent where the TRIS molecules induce an osmotic pressure in the microcapsules' interior. This pressure drives the inflation of the LbL microcapsules that causes the expansion of the LbL capsular walls. Fluorescence staining reveals that the inflated LbL microcapsules consist of an agarose microbead suspended within the aqueous interior of the capsule but still attached to the LbL capsular wall at one point; thereby displaying a ‘bead-in-a-capsule’ morphology. It was demonstrated that the degree of inflation depends on the concentration of pre-loaded TRIS and the number of coated polymer layers. Also, ADOGEN^® 464 (a cationic surfactant) is required for the fabrication of the inflated LbL microcapsules. The mass of dextran macromolecules (65–2000 kDa) diffusing through the LbL capsular wall had decreased by at least 49% after expansion of the capsular wall. Inflated microcapsules were shown to be capable of controlling the distribution of two different materials internally. Hence, it is possible that inflated microcapsules can permit localized control over chemical or enzymatic reactions for future uses in biomedical applications.

We report on a novel method for the encapsulation of highly water soluble materials by using layer-by-layer (LbL) polyelectrolyte self-assembly. State of the art polyelectrolyte self-assembly LbL coating and encapsulation methods are only applicable to insoluble or poorly water soluble template materials, because the process is performed in water causing dissolution of the solid template. Our method extends the material spectrum to highly water soluble template materials by using non-ionized polyelectrolytes in an organic phase (reverse-phase) instead of polyelectrolyte salts in an aqueous environment. By using the reverse-phase layer-by-layer (RP-LbL) technique, we have demonstrated the direct encapsulation of proteins, glucose, vitamin C, and inorganic salts in the solid state. Multilayer deposition was proven, layer thickness was determined by AFM, and the advantage of the method to prepare powders of encapsulated materials was demonstrated. The method is simple, robust, and applicable to a broad range of substances with potential applications in several industries.

Point-of-care (POC) diagnostics brings tests nearer to the site of patient care. The turnaround time is short, and minimal manual interference enables quick clinical management decisions. Growth in POC diagnostics is being continuously fueled by the global burden of cardiovascular and infectious diseases. Early diagnosis and rapid initiation of treatment are crucial in the management of such patients. This review provides the rationale for the use of POC tests in acute coronary syndrome, heart failure, human immunodeficiency virus, and tuberculosis. We also consider emerging technologies that are based on advanced nanomaterials and microfluidics, improved assay sensitivity, miniaturization in device design, reduced costs, and high-throughput multiplex detection, all of which may shape the future development of POC diagnostics.

Polypyrrole magnetic microspheres were synthesized and used to extract carbaryl, carbofuran, and methomyl before analysis by a high-performance liquid chromatography with diode array detection. Under optimal conditions, four times the preconcentration was achieved with the use of only 1.2 mL of sample. Good linearity with ranges of 3.0–7.5 × 103, 6.0–4.5 × 103, and 15–3.0 × 103 ng kg−1 and limits of detection of 1.37 ± 0.10, 4.7 ± 1.2, and 10.1 ± 5.7 ng kg−1 were obtained, respectively. Good reproducibility (RSDs < 5%) was achieved over 24 cycles of extraction and regeneration. Good accuracy (recoveries 81.6 ± 1.5%–108.3 ± 2.2%) and good precision (RSDs 0.11%–4.5%) were obtained. Carbaryl was detected in apple (2.75 ± 0.23 ng kg−1), carbofuran in tomato (11.34 ± 0.61 ng kg−1), and methomyl in watermelon (34.7 ± 1.7 ng kg−1). The relative expanded uncertainty of the measurement method was less than 14% for all three pesticides.

Abstract Lignosulfonate (LS) is a large-scale surplus product of the forest and paper industries, and has primarily been utilized as a low-cost plasticizer in making concrete for the construction industry. LS is an anionic redox-active polyelectrolyte and is a promising candidate to boost the charge capacity of the positive electrode (positrode) in redox-supercapacitors. Here, the physical-chemical investigation of how this biopolymer incorporates into the conducting polymer PEDOT matrix, of the positrode, by means of counter-ion exchange is reported. Upon successful incorporation, an optimal access to redox moieties is achieved, which provides a 63% increase of the resulting stored electrical charge by reversible redox interconversion. The effects of pH, ionic strength, and concentrations, of included components, on the polymer?polymer interactions are optimized to exploit the biopolymer-associated redox currents. Further, the explored LS-conducting polymer incorporation strategy, via aqueous synthesis, is evaluated in an up-scaling effort toward large-scale electrical energy storage technology. By using an up-scaled production protocol, integration of the biopolymer within the conducting polymer matrix by counter-ion exchange is confirmed and the PEDOT-LS synthesized through optimized strategy reaches an improved charge capacity of 44.6 mAh g?1.

Microparticles have attracted much attention for medical, analytical and biological applications. Calcium carbonate (CaCO3) templating method with the advantages of having narrow size distribution, controlled morphology and good biocompatibility that has been widely used for the synthesis of various protein-based microparticles. Despite CaCO3 template is biocompatible, most of the conventional methods to create stable protein microparticles are mainly driven by chemical crosslink reagents which may induce potential harmful effect and remains undesirable especially for biomedical or clinical applications. In this article, we demonstrate the fabrication of protein microparticles and microcapsules with an innovative method using biomolecular tools such as enzymes and affinity molecules to trigger the assembling of protein molecules within a porous CaCO3 template followed by a template removal step. We demonstrated the enzyme-assisted fabrication of collagen microparticles triggered by transglutaminase, as well as the affinity-assisted fabrication of BSA-biotin avidin microcapsules triggered by biotin-avidin affinity interaction, respectively. Based on the different protein assemble mechanisms, the collagen microparticles appeared as a solid-structured particles, while the BSA-biotin avidin microcapsules appeared as hollow-structured morphology. The fabrication procedures are simple and robust that allows producing protein microparticles or microcapsules under mild conditions at physiological pH and temperature. In addition, the microparticle morphologies, protein compositions and the assemble mechanisms were studied. Our technology provides a facile approach to design and fabricate protein microparticles and microcapsules that are useful in the area of biomaterials, pharmaceuticals and analytical chemistry.

A reusable optical bioassay platform using permeability-controlled hydrogel pads for selective saccharide detection has been developed. An optical glucose detection assay based on fluorescence resonance energy transfer (FRET) between dye-labeled dextran and Concanavalin A (ConA) was incorporated into hydrogel pads by entrapment. The hydrogel pads are constructed from hemispherical hydrogel attached onto hydrophobic surfaces of a microtiter plate. The resulted hemispherical hydrogel pads entrapping the sensing biological materials were further surface coated with polyelectrolyte multilayers through a Layer-by-Layer (LbL) self-assembly process to create a permeability-controlled membrane with nanometer thickness. The selective permeable LbL film deposited on the hydrogel surface allows small molecular weight analytes to diffuse into the hydrogel pads while the large molecular weight sensing biological molecules are immobilized. An encapsulation efficiency of 75% for the ConA/Dextran complex within the coated hydrogel pads was achieved and no significant leakage of the complex was observed. Glucose calibration curve with linear range from 0 to 10 mM glucose was obtained. Selective permeability of the hydrogel pads has been demonstrated by measurement of saccharides with various molecular weights. The LbL hydrogel pads could selectively detect monosaccharides (glucose, MW = 180) and disaccharides (sucrose, MW = 342) while polysaccharides (dextran, MW ∼ 70 kDa) cannot diffuse through the LbL layer and are excluded. LbL hydrogel pads allow regeneration of the FRET system with good signal reproducibility of more than 90% to construct a reusable and reagentless optical bioassay platform.

We have demonstrated an entirely new concept of a wearable theranostic device in the form of a contact lens (theranostic lens) with a dual-functional hybrid surface to modulate and detect a pathogenic attack, using a the corneal HSV serotype-1 (HSV-1) model. The theranostic lenses were constructed using a facile layer-by-layer surface engineering technique, keeping the theranostic lenses with good surface wettability, optically transparency, and nontoxic toward human corneal epithelial cells. The theranostic lenses were used to capture and concentrate inflammatory cytokines such as interleukin-1 alpha (IL-1 alpha), which is upregulated during HSV-1 reactivation, for sensitive, noninvasive diagnostics. The theranostic lens also incorporated an antiviral coating to serve as a first line of defense to protect patients against disease. Our strategy tackles major problems in tear diagnostics that are mainly associated with the sampling of a relatively small volume of fluid and the low concentration of biomarkers. The theranostic lenses show effective anti-HSV-1 activity and good analytical performance for the detection of IL-1a, with a limit of detection of 1.43 pg mL(-1) and a wide linear range covering the clinically relevant region. This work offers a new paradigm for wearable noninvasive healthcare devices combining diagnosis and protection against disease, while supporting patient compliance. We believe that this approach holds immense promise as a next-generation point-of-care and decentralized diagnostic/theranostic platform for a range of biomarkers.

The nitrocellulose (NC) membrane based lateral flow immunoassay device (LFID) is one of the most important and widely used biosensor platforms for point-of-care (PoC) diagnostics. However, the analytical performance of LFID has limitations and its optimization is restricted to the bioassay chemistry, the membrane porosity, and the choice of biolabel system. These bottom neck technical issues resulted from the fact that the conventional LFID design principle has not evolved for many years, which limited the LFID for advanced biosensor applications. Here we introduce a new dimension for LFID design and optimization based on geometric flow control (GFC) of NC membranes, leading to highly sensitive GFC-LFID. This novel approach enables comprehensive flow control via different membrane geometric features such as the width (w) and the length (l) of a constriction, as well as its input angle (?1) and output angle (?2). The GFC-LFID (w=0.5 mm, l=7 mm, ?1= 60, ?2= 45) attained a 10-fold increase in sensitivity for detection of interleukin-6 (IL-6), compared with conventional LFID, whereas reducing by 10-fold the antibody consumption. The GFC-LFID detects IL-6 over a linear range of 0.1-10?ng/mL with a limit of detection (LoD) of 29?pg/mL, which even outperforms some commercial IL-6 LFIDs. Such significant improvement is attained by pure geometric control of the NC membrane, without additives, that only relaying on a simple high throughput laser ablation procedure suitable for integration on regular large-scale manufacturing of GFC-LFIDs. Our new development on GFC-LFID with the combination of facile scalable fabrication process, tailored flow control, improved analytical performance, and reduced antibodies consumption is likely to have a significant impact on new design concept for the LFID industry.

We have demonstrated a new approach using a neutravidin-based biosensor combined with a dual-function gold nanoparticle (AuNP) biolabel, for simple and sensitive detection of microRNA-21 (miRNA-21). The selectivity of the biosensor is provided by the intrinsic properties of the dual-functional biotin-MB-AuNP label. The assay procedure is relatively simple, exploiting a one-pot assay concept where the affinity capture of the miRNA-21/dual-functional biotin-MB-AuNP complex, via the strong biotin-neutravidin supramolecular interaction, and simultaneous detection of the captured AuNPs label with stripping voltammetry, is performed in a single step. This electrochemical miRNA biosensor could detect miRNA-21 with limit of detection of 0.1×10less thansuperscriptgreater than−12less than/superscriptgreater than and a dynamic range from 0.5×10less thansuperscriptgreater than−12less than/superscriptgreater than to 1.0×10less thansuperscriptgreater than−9less than/superscriptgreater thanM. The performance of the miRNA-21biosensor was further improved after silver deposition onto the AuNPs, delivering an enhanced detection limit of 4.0×10less thansuperscriptgreater than−15less than/superscriptgreater thanM of miRNA-21, and an extremely wide analytic dynamic range from 10×10less thansuperscriptgreater than−15less than/superscriptgreater than to 1×10less thansuperscriptgreater than−9less than/superscriptgreater thanM (5 orders of magnitude). This exceptionally broad dynamic range demonstrates the advantage of the one-pot assay approach with direct capture of the dual functional biotin-MB-AuNP via the strong biotin-neutravidin supramolecular interaction. Furthermore, we demonstrated the detection of miRNA-21 in spiked serum at clinically relevant concentrations. The miRNA biosensor displayed excellent analytical performance for the detection of miRNA and could provide a powerful and convenient tool for biomedical research and applications in cancer diagnostics.

Natural polymer-based nanofibrous structures promote cell adhesion and proliferation due to their high surface area/volume ratio, high porosity, and similarity to native extracellular matrix in terms of both chemical composition and physical structure. Gum tragacanth (Tg) is a natural polysaccharides obtained from plants. It is a biocompatible, biodegradable and anionic polysaccharides that has been used extensively as an emulsifier in food and pharmaceutical industries. Despite, its good rheological properties and compatibility, the potential biomedical applications of Tg have not been fully investigated. The objective of the present study was to explore the feasibility of combining Tg with gelatin to fabricate a scaffold that serves as a simple collagen-glycosaminoglycans analog for tissue engineering applications, e.g. as a scaffold for human skin epithelial cells.

Recently, one-dimensional nanostructures with different morphologies (such as nanowires, nanorods (NRs), and nanotubes) have become the focus of intensive research, because of their unique properties with potential applications. Among them, zinc oxide (ZnO) nanomaterials has been found to be highly attractive, because of the remarkable potential for applications in many different areas such as solar cells, sensors, piezoelectric devices, photodiode devices, sun screens, antireflection coatings, and photocatalysis. Here, we present an innovative approach to create a new modified textile by direct in situ growth of vertically aligned one-dimensional (1D) ZnO NRs onto textile surfaces, which can serve with potential for biosensing, photocatalysis, and antibacterial applications. ZnO NRs were grown by using a simple aqueous chemical growth method. Results from analyses such as X-ray diffraction (XRD) and scanning electron microscopy (SEM) revealed that the ZnO NRs were dispersed over the entire surface of the textile. We have demonstrated the following applications of these multifunctional textiles: (1) as a flexible working electrode for the detection of aldicarb (ALD) pesticide, (2) as a photo catalyst for the degradation of organic molecules (i.e., Methylene Blue and Congo Red), and (3) as antibacterial agents against Escherichia coli. The ZnO-based textile exhibited excellent photocatalytic and antibacterial activities, and it showed a promising sensing response. The combination of sensing, photo catalysis, and antibacterial properties provided by the ZnO NRs brings us closer to the concept of smart textiles for wearable sensing without a deodorant and antibacterial control. Perhaps the best known of the products that is available in markets for such purposes are textiles with silver nanoparticles. Our modified textile is thus providing acceptable antibacterial properties, compared to available commercial modified textiles.

Single and binary component patterning and integrated microfabrication of biomolecules, such as DNA and proteins, can be achieved by focused ion-beam (FIB) biolithography. Well-defined micropatterns are obtained by FIB milling on biomolecules immobilized on SiO₂ wafers and protected by a thin Au film. The retention of biofunctionality is excellent (68–90%) and a feature size of down to 500 nm can be achieved for the patterns without significant loss of functionality.

Encoding by encapsulation: A polymeric shell fabrication approach combines biomolecule encapsulation with encoding. Striated polymeric shells, fabricated through an inwards diffusion of poly(allylamine) into the matrices of agarose microbeads, serves to encapsulate the biomolecules within the microcapsule. Encoding is performed through the color and/or thickness permutation of the striated polymeric shells (see picture).

The oxygen reduction reaction (ORR) is an essential process in electrocatalysis limiting the commercialization of sustainable energy conversion technologies, such as fuel cells. The use of conducting polymers as molecular porous and conducting catalysts obtained from the high abundance elements enables the route towards low cost and high-throughput fabrication of disposable plastic electrodes of fuel cells. Poly(3,4-ethylenedioxythiophene) (PEDOT) is a 2-electron ORR electrocatalyst yielding specifically hydrogen peroxide that limits the full utilization of chemical energy of oxygen. Here, we demonstrated an innovative product-to-intermediate relay approach achieving complete oxygen reduction reaction (cORR) with Prussian blue (PB) integrated microporous PEDOT cathode in fuel cells. The microporous structured PEDOT electrode prepared via a simple cryosynthesis allows the bulk integration and stabilization of the poor conducting PB co-catalyst into the PEDOT ion-electron conductor, while the microporous PEDOT allows effective oxygen diffusion into the matrix. We evaluated systematically the effect of sequential PEDOT 2-electron ORR followed by PB co-catalysis launching hydrogen peroxide reduction reaction (HPRR) into H2O. This resulted in the establishment of electronic and ionic transport between PEDOT and PB catalyst enabling the combination of enhanced ORR electrocatalysis by means of the ORR course extension from 2to 4-electron reduction to achieve cORR. The cORR performance delivered by the product-to-intermediate relay between microporous PEDOT and PB co-catalysis led to a four times increase in power density of model proton-exchange membrane fuel cell (PEMFC) assembled from the polymer-based air breathing cathode.

In this article, we report an innovative green chemistry approach for the fabrication of protein microparticles based on peptide mediated disulfide interchange reactions. The concept is based on using a redox reactive peptide, glutathione, as a natural crosslink reagent triggering the formation of intermolecular disulfide bonds between adjacent protein molecules leading to the assembly of protein molecules within a CaCO3 template into a microparticle structure. The CaCO3 template is highly biocompatible and is completely removed by simply adjusting the solution to pH 5.0, leaving behind the pure protein microparticles. Moreover, the GSH is only involved in the intermediate step without being incorporated into the resulting protein microparticles, therefore producing protein microparticles composed of purely protein molecules. This technology provides a simple and robust method to fabricate protein microparticles under physiological aqueous conditions, and more importantly avoiding the extensive use of synthetic chemical crosslinking reagents. We have further demonstrated that this method is versatile to fabricate microparticles with various proteins such as BSA, enzymes and antibodies. The biological functions such as catalytic properties and affinity interactions of the resulting protein microparticles are highly conserved which demonstrate the potential applications of the protein microparticles in the area of biocatalysis, bioseparation and targeted drug delivery.

Flexible electronics have gained more attention for emerging electronic devices such as sensors, biosensors, and batteries with advantageous properties including being thin, lightweight, flexible, and low-cost. The development of various forms of allotropic carbon papers provided a new dry-manufacturing route for the fabrication of flexible and wearable electronics, while the electrochemical performance and the bending stability are largely influenced by the bulk morphology and the micro-/nanostructured domains of the carbon papers. Here, we evaluate systematically the intrinsic physicoelectrochemical properties of allotropic carbon-based conducting papers as flexible electrodes including carbon-nanotubes-paper (CNTs-paper), graphene-paper (GR-paper), and carbon-fiber-paper (CF-paper), followed by functionalization of the allotropic carbon papers for the fabrication of flexible electrodes. The morphology, chemical structure, and defects originating from the allotropic nanostructured carbon materials were characterized by scanning electron microscopy (SEM) and Raman spectroscopy, followed by evaluating the electrochemical performance of the corresponding flexible electrodes by cyclic voltammetry and electrochemical impedance spectroscopy. The electron-transfer rate constants of the CNTs-paper and GR-paper electrodes were similar to 14 times higher compared with the CF-paper electrode. The CNTs-paper and GR-paper electrodes composed of nanostructured carbon showed significantly higher bending stabilities of 5.61 and 4.96 times compared with the CF-paper. The carbon-paper flexible electrodes were further functionalized with an inorganic catalyst, Prussian blue (PB), forming the PB-carbon-paper catalytic electrode and an organic conducting polymer, poly(3,4-ethylenedioxythiophene) (PEDOT), forming the PEDOT-carbon-paper capacitive electrode. The intrinsic attribute of different allotropic carbon electrodes affects the deposition of PB and PEDOT, leading to different electrocatalytic and capacitive performances. These findings are insightful for the future development and fabrication of advanced flexible electronics with allotropic carbon papers.

An innovative modular approach for facile design and construction of flexible microfluidic biosensor platforms based on a dry manufacturing "craft-and-stick" approach is developed. The design and fabrication of the flexible graphene paper electrode (GPE) unit and polyethylene tetraphthalate sheet (PET)6/adhesive fluidic unit are completed by an economic and generic xurographic craft approach. The GPE widths and the microfluidic channels can be constructed down to 300 mu m and 200 mu m, respectively. Both units were assembled by simple double-sided adhesive tapes into a microfluidic integrated GPE (MF-iGPE) that are flexible, thin (&lt;0.5 mm), and lightweight (0.4 g). We further functionalized the iGPE with Prussian blue and glucose oxidase for the fabrication of MF-iGPE glucose biosensors. With a closed-channel PET fluidic pattern, the MF-iGPE glucose biosensors were packaged and sealed to protect the integrated device from moisture for storage and could easily open with scissors for sample loading. Our glucose biosensors showed 2 linear dynamic regions of 0.05-1.0 and 1.0-5.5 mmol L-1 glucose. The MF-iGPE showed good reproducibility for glucose detection (RSD &lt; 6.1%, n = 6) and required only 10 mu L of the analyte. This modular craft-and-stick manufacturing approach could potentially further develop along the concept of paper-crafted model assembly kits suitable for low-resource laboratories or classroom settings.

A single-probe strip test for the rapid and sensitive detection of miRNA-21 mimics is reported herein. Highly specific structurally responsive bi-functional, thiol and biotin, DNA/LNA oligonucleotide probes (molecular beacons-MB) were designed and conjugated with gold nanoparticles (AuNPs) (i.e. biotin-MB-AuNPs). The proposed design had the ability to modulate the accessibility of the biotin group as a function of the presence of a miRNA target allowing the interaction of the boilable with the streptavidin test zone only in the presence of the miRNA-21 mimics. For quantitative evaluation, images of the strip tests were recorded using a flatbed scanner (Epson Perfection V370 Photo). The colour intensities of the test zones of the strip tests were analysed with the ImageJ software (Scion Corp., USA) and quantified as a function of pixel intensity. The response of the strip test was linear over the range 0.5 to 20 nM miRNA-21 (limit of detection of 115 pM) and showed good reproducibility (intra and inter CVs below 8 %); furthermore, the assay was shown to be highly selective, discriminating other interference miRNAs mimics (e.g. miRNA-221 and miRNA-205). Finally, the proposed strip test was used for detection of miRNA-21 mimics in spiked serum samples, demonstrating its potential for point-of-care clinical applications. Main advantages of the single-probe strip test design are its versatility, simplicity and robustness, which can be easily extended to other miRNA targets by tuning the sequence of the single probe. Furthermore, the use of the structurally responsive single probe is particularly relevant in the case of short-length targets, such as miRNA, whereas a conventional sandwich approach might require a careful control of assay conditions such as hybridization temperature and salt concentration

Recent advances in the field of dermatotherapy have resulted in research efforts focusing on the use of particle-based drug delivery systems for the stimuli-responsive release of drugs in the skin and skin appendages, i.e. hair follicles and sebaceous glands. However, effective and innocuous trigger mechanisms which result in the release of the drugs from the nanocarriers upon reaching the target structures are still lacking. For the first time, the present study demonstrated the photo-activated release of the model drug fluorescein isothiocyanate (FITC) from topically applied gold nanoparticle-doped bovine serum albumin (AuNPs-doped BSA) particles (approx. 545 nm) using water-filtered infrared A (IRA) radiation in the hair follicles of an ex vivo porcine skin model. The IRA radiation-induced plasmonic heating of the AuNPs results in the partial decomposition or opening of the albumin particles and release the model drug, while control particles without AuNPs show insignificant release. The results demonstrate the feasibility of using IRA radiation to induce release of encapsulated drugs from plasmonic nanocarriers for the targeting of follicular structures. However, the risk of radiation-induced skin damage subsequent to repeated applications of high infrared dosages may be significant. Future studies should aim at determining the suitability of lower infrared A dosages, such as for medical treatment regimens which may necessitate repeated exposure to therapeutics. Statement of significance Follicular targeting using nanocarriers is of increasing importance in the prophylaxis and treatment of dermatological or other diseases. For the first time, the present study demonstrated the photo activated release of the model drug fluorescein isothiocyanate (FITC) from topically applied gold nanoparticle-doped bovine serum albumin (AuNPs-doped BSA) particles using water-filtered infrared A (IRA) radiation in the hair follicles of an ex vivo porcine skin model. The results demonstrate the feasibility of using wIRA radiation to induce release of encapsulated drugs for the targeting of follicular structures, and provide a new vision on the development of optically addressable delivery systems for controlled release of drugs in the skin and skin appendages, i.e. hair follicles and sebaceous glands. (C) 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

The efficacy of topically applied drugs is determined by their action mechanism and their potential capacity of passing the skin barrier. Nanoparticles are assumed to be efficient carrier systems for drug delivery through the skin barrier. For flexible nanoparticles like liposomes, this effect has been well demonstrated. The penetration properties of solid nanoparticles are currently under intensive investigation. The crucial advantage of nanoparticles over non-particulate substances is their capability to penetrate deeply into the hair follicles where they can be stored for several days. There is no evidence, yet, that solid particles ≥40 nm are capable of passing through the healthy skin barrier. Therefore and in spite of the long-standing research efforts in this field, commercially available solid nanoparticle-based products for drug delivery through the healthy skin are still missing. Nevertheless, the prospects for the clinical use of nanoparticles in drug delivery are tremendous. They can be designed as transport systems delivering drugs efficiently into the hair follicles in the vicinity of specific target structures. Once deposited at these structures, specific signals might trigger the release of the drugs and exert their effects on the target cells. In this article, examples of such triggered drug release are presented.

Biomaterial-based microparticles have attracted much attention for medical and biological applications such as pharmaceutics, bioseparation and cosmetics. Emerging technologies enable versatile and facile fabrication of microparticles, with key features being purity, precise size control, mild preparation conditions and minimal processing. Here, an innovative approach combining template synthesis, biomolecule assembly and partial-purification within a single step for high efficiency fabrication of pure biomaterial-based microparticles is reported. This concept is based on facile co-precipitation of biomolecules within CaCO₃ templates and simultaneous crosslinking of entrapped biomolecules via Ca²⁺ driven supramolecular coordination chemistry, followed by template removal. Carbohydrate (alginate) and proteins (casein and fresh milk) are used as models of biomolecules. The process driven by selective crosslinking automatically excludes non-specific materials from the template and thus provides the additional function of partial-purification, as demonstrated using highly complexed fresh milk. This green approach to fabrication of biomaterial-based microparticles offers three critical advantages (i) mild conditions to preserve the chemical and secondary structures of biomolecules; (ii) single processing step to facilitate scale-up production; and (iii) partial-purification without the need for upstream raw material purification. This innovative approach not only addresses fundamental issues in fabrication techniques, but also marks progress in energy and environmental conservation during manufacturing processes.

A new approach for the facile fabrication of electrochemical biosensors using a biohybrid conducting polymer was demonstrated using glucose oxidase (GOx) and poly (3, 4-ethylenedioxythiophene) (PEDOT) as a model. The biohybrid conducting polymer was prepared based on a template-assisted chemical polymerisation leading to the formation of PEDOT microspheres (PEDOT-MSs), followed by in-situ deposition of platinum nanoparticles (PtNPs) and electrostatic immobilisation of glucose oxidase (GOx) to form water processable GOx-PtNPs-PEDOT-MSs. The morphology, chemical composition and electrochemical performance of the GOx-PtNPs-PEDOT-MS-based glucose biosensor were characterised using scanning electron microscopy (SEM), energy-dispersive X-ray spectrometry (EDS), Fourier transform infrared (FTIR) spectroscopy, zeta potential and electrochemical measurements, respectively. The biosensor delivered a linear response for glucose over the range 0.1-10 mM (R-2 = 0.9855) with a sensitivity of 116.25 mu A mM(-1) cm(-2), and limit of detection of 1.55 mu M (3 x SD/sensitivity). The sensitivity of the developed PEDOT-MS based biosensor is significantly higher (2.7 times) than the best reported PEDOT-based glucose biosensor in the literature. The apparent Michaelis Menten constant (K-m(app)) of the GOx-PtNPs-PEDOT-MS-based biosensors was calculated as 7.3 mM. Moreover, the biosensor exhibited good storage stability, retaining 97% of its sensitivity after 12 days storage. This new bio-hybrid conducting polymer combines the advantages of micro-structured morphology, compatibility with large-scale manufacturing processes, and intrinsic biocatalytic activity and conductivity, thus demonstrating its potential as a convenient material for printed bioelectronics and sensors.

Much recent work has focused on improving the processibility and electrocapacitive performance of conducting polymer-based materials for energy related applications. The key mechanism of conducting polymers as supercapacitor materials is driven by the rapid charging and discharging processes that involve mass transport of the counter ions insertion/ejection within the polymer structure, where ion diffusion is usually the limiting step on the efficiency of the conducting polymer capacitor. Here, we report a facile method for the green fabrication of polypyrrole microspheres (PPy-MSs) and poly (3, 4-ethylenedioxythiophene) microspheres (PEDOT-MSs) with good processability, intact morphology and large active surface for enhanced ion interchange processes, without using surfactant and highly irritant or toxic organic solvents during the synthetic process. The structure and morphology of the PPy-MSs and PEDOT-MSs were characterized by means of SEM, EDX, TEM and FTIR. Both PPy-MSs and PEDOT-MSs showed intact microsphere structures with greatly improved water dispersity and processability. More importantly, facilated by the large active surface and inter-microsphere space for ions diffusion, both the PPy-MSs and PEDOT-MSs showed a signiciantly enhanced electrical capacitive performance of 242 F g(-1) and 91.2 F g(-1), repsectively (i.e. 10 and 1.51 times in specific capacitance than the randomly structured PPy and PEDOT). This innovative approach not only addresses fundamental issues in fabrication of high performance processable microstructured conducting polymers, but also makes progress in delivering water processable conducting polymers that could be potentially used for fabrication of printed electronic devices.

Integrated printed microfluidic biosensors are one of the most recent point-of-care (POC) sensor developments. Fast turnaround time for production and ease of customization, enabled by the integration of recognition elements and transducers, are key for on-site biosensing for both healthcare and industry and for speeding up translation to real-life applications. Here, we provide an overview of recent progress in printed microfluidics, from the 2D to the 4D level, accompanied by novel sensing element integration. We also explore the latest trends in integrated printed microfluidics for healthcare, especially POC diagnostics, and food safety applications.

This paper presents the general principles of various biosensor systems, reviews current biosensor technologies for agricultural and environmental monitoring, and discusses their opportunities and challenges. Advances in biosensor technologies could provide a useful analytical tools for agricultural monitoring, particularly due to their rapid response, relatively low operational cost and portability for field/farm application. The promise, demonstrated by various examples of biosensor technologies, is very appealing. However, there are still many hurdles to bring commercial agricultural biosensors into real practice.

The layer-by-layer (LbL) polyelectrolyte self-assembly encapsulation method has attracted much interest because of its versatility to use various polymers for capsule formation, ability to encapsulate different templates, and capability to control capsule permeability. Traditionally, the LbL method was performed in water as solvent and limited to poorly or non-water-soluble templates. Using the matrix-assisted LbL method, complex mixtures of water-soluble proteins or DNA could be encapsulated within agarose microbeads templates but leakage of biomolecules into the water phase during the LbL process results in low encapsulation efficiency. Recently, the reverse-phase LbL (RP-LbL) method was introduced to perform LbL and encapsulation of water-soluble templates in organic solvents, thus preventing the templates from dissolving and allowing high encapsulation efficiency. However, encapsulation of complex mixtures of biomolecules or other substances with quantitative encapsulation efficiency remained impossible. Here we present a new approach for encapsulation of biomolecules or complex mixtures thereof with almost 100% encapsulation efficiency. The ability of our method to achieve high encapsulation efficiency arises from the combination of two strategies. (1) Using microparticles as surface stabilizer to create stable biomolecule-loaded hydrogel microbeads, termed matrix-assisted colloidosome (MAC), that are able to disperse in oil and organic solvents. (2) Using the RP-LbL method to fabricate polymeric capsule “membranes”, thereby preventing diffusion of the highly water-soluble biomolecules. Using an oil phase during emulsification and an organic solvent phase during encapsulation could completely prevent leakage of water-soluble biomolecules and almost 100% encapsulation efficiency is achieved. Microcapsules fabricated with our method retained nearly 100% of encapsulated proteins during a 7 day incubation period in water. The method was demonstrated on model proteins and may be extended to other biomolecules or mixtures. Our method is a valuable addition to the family of encapsulation techniques and can significantly contribute to the fields of bioreactors and bioanalytical microcapsules.

Lateral-flow tests were first launched commercially in 1984, as a simple urine-based pregnancy test for home use. The simplicity of the visual readout delivered by the basic lateral-flow format proved to be a very popular. However, the recent apparently unstoppable trend towards portable and wearable technology is driving the lateral-flow strip towards an industrial interface that will enable it to interface with big data and expert systems, and where ready transmission of data is essential. In this review, we chart the inevitable evolution of the visually-read lateral-flow strip to more advanced instrumented versions and consider the future of this very flexible approach to delivering simple affinity assays. We examine recent labelling strategies, the relative merits of optical and electrochemical transducers and explore the evolution of recognition elements that are now being incorporated into these systems.

A thick-film phosphate biosensor based on hydrogel immobilized pyruvate oxidase (POD) has been developed for rapid phosphate process control monitoring in an experimental sequencing batch reactor (SBR) system. We have employed a phosphate biosensor in an off-line monitoring of phosphate concentrations in a bench scale SBR. Measurements with biosensor show a good correlation (r2=0.98) with those of commercial colorimetric phosphate testing kits. The signal response time was 1 min with a detection limit of 5 microM. The biosensor method showed a good operational stability, needed less experimental procedures and a small sample size (approximately 20 microl). This allows its practical application for rapid phosphate measurements to obtain real time process data in a SBR system.

A novel approach to perform a high number of individual polymerase chain reactions (PCR) in microcapsule reaction compartments, termed “Microcapsule-PCR” was developed. Temperature stable microcapsules with a selective permeable capsule wall were constructed by matrix-assisted layer-by-layer (LbL) Encapsulation technique. During the PCR, small molecular weight building blocks – nucleotides (dNTPs) were supplied externally and diffuse through the permeable capsule wall into the interior, while the resulted high molecular weight PCR products were accumulated within the microcapsule. Microcapsules (∼110.8 µm average diameter) filled with a PCR reaction mixture were constructed by an emulsion technique having a 2% agarose core and a capsule formed by LbL coating with poly(allylamine-hydrochloride) and poly(4-styrene-sulfonate). An encapsulation efficiency of 47% (measured for primer-FITC (22 bases)) and 98% PCR efficiency was achieved. Microcapsules formed by eight layers of polyelectrolyte and subjected to PCR cycling (up to 95 °C) demonstrated good temperature stability without any significantly changes in DNA retention yield and microcapsule morphology. A multiplex Microcapsule-PCR experiment demonstrated that microcapsules are individual compartment and do not exchange templates or primers between microcapsules during PCR cycling.

A new approach to perform electrochemical immunoassay based on the utilization of encapsulated microcrystal was developed. The microcrystal labels create a “supernova effect” upon exposure to a desired releasing agent. The microcrystal cores dissolve, and large amounts of signal-generating molecules diffuse across the capsule wall into the outer environment. Layer-by-Layer (LbL) technology was employed for the encapsulation of electrochemical signal-generating microcrystals (ferrocene microcrystals). The encapsulated microcrystals were conjugated with antibody molecules through the adsorption process. The biofunctionalized microcrystals were utilized as a probe for immunoassays. The microcrystal-based label system provided a high-signal molecule to antibody (S/P) ratio of 10⁴−10⁵. Microcrystal biolabels with different antibody surface coverage (1.60−5.05 mg m^-2) were subjected to a solid-phase immunoassay for the detection of mouse immunoglobulin G (M-IgG) molecules. The microcrystal-based immunoassay for the detection of M-IgG performed with microcrystals having antibody surface coverage of 5.05 mg m^-2 showed a sensitivity of 3.93 nA μg^-1 L^-1 with a detection limit of 2.82 μg L^-1.

In this article, a non-chemical crosslinking method is used to produce pure protein microparticles with an innovative approach, so-called protein activation spontaneous and self-assembly (PASS). The fabrication of protein microparticles is based on the idea of using the internal disulfide bridges within protein molecules as molecular linkers to assemble protein molecules into a microparticle form. The assembly process is triggered by an activating reagent–dithiothreitol (DTT), which only involved in the intermediate step without being incorporated into the resulting protein microparticles. Conventional protein microparticle fabrication methods usually involve emulsification process and chemical crosslink reactions using amine reactive reagents such as glutaraldehdye or EDC/NHS. The resulting protein microparticles are usually having various size distributions. Most importantly crosslinking reactions using amine reactive reagents will result in producing protein microparticles with undesired properties such as auto-fluorescence and high toxicity. In contrast to the conventional methods, our technology provides a simple and robust method to produce highly homogeneous, stable and non-fluorescence pure protein microparticles under mild conditions at physiological pH and temperature. The protein microparticles are found to be biodegradable, non-toxic to MDCK cells and with preserved biological activities. Results on the cytotoxcity study and enzyme function demonstrate the potential applications of the protein microparticles in the area of pharmaceutics and analytical chemistry.

We report on the nanoengineered encapsulation and functionalization of mediator microcrystals and their use as a non-metallic bioassay label system for the silver enhancement technology. Mediator microcrystals were encapsulated and functionalized by using a Layer-by-Layer (LbL) based approach. This is a generic way to transform most low-soluble mediators (e.g., ferroceneacetic acid) into a label for electrochemical bioassays. Instead of using conventional gold particles, encapsulated mediator microcrystals are used as the primary label for the silver enhancement technique. After the biorecognition reaction the unbound labels were separated and a silver(I) ion solution was added. The mediator reduces silver ions into metallic silver and the mediator microcrystal was transformed into a silver particle. Consecutively a reducing agent was added leading to a further silver growth, representing the silver enhancement step. A large amount of silver was developed from the mediator, resulting in signal amplification. The amount of developed silver was studied in a bioassay and was found proportional to the analyte concentration.

Human mesenchymal stromal cells (hMSCs) are adult-source cells that have been extensively evaluated for cell-based therapies. hMSCs delivered by intravascular injection have been reported to accumulate at the sites of injury to promote tissue repair and can also be employed as vectors for the delivery of therapeutic genes. However, the full potential of hMSCs remains limited as the cells are lost after injection due to anoikis and the adverse pathologic environment. Encapsulation of cells has been proposed as a means of increasing cell viability. However, controlling the release of therapeutic cells over time to target tissue still remains a challenge today. Here, we report the design and development of thermo-rheological responsive hydrogels that allow for precise, time dependent controlled-release of hMSCs. The encapsulated hMSCs retained good viability from 76% to 87% dependent upon the hydrogel compositions. We demonstrated the design of different blended hydrogel composites with modulated strength (S parameter) and looseness of hydrogel networks (N parameter) to control the release of hMSCs from thermoresponsive hydrogel capsules. We further showed the feasibility for controlled-release of encapsulated hMSCs within 3D matrix scaffolds. We reported for the first time by a systematic analysis that there is a direct correlation between the thermo-rheological properties associated with the degradation of the hydrogel composite and the cell release kinetics. This work therefore provides new insights into the further development of smart carrier systems for stem cell therapy.

Phytase (EC 3.1.3.26) and phytic acid (myo-inositol hexaphosphate) play an important environmental role in poultry industry and have a health aspect in food industry. Novel biosensors have been developed for simple, one step quantitative phytic acid and phytase detection. A system based on the sequentially acting enzyme phytase and pyruvate oxidase (POD) was employed for the development of phytase and phytic acid biosensors. Poly(carbamoylsulphonate) (PCS) hydrogel immobilized POD electrode was applied for the detection of phytase. It was based on the indication of phosphate ions produced by the hydrolysis of phytic acid. The phytase biosensor showed a linear response ranging from 0.5 to 6.0 units/ml. A bi-enzyme sensor based on co-immobilization of phytase and POD was developed for the detection of phytic acid on the basis of amperometric detection of the enzymatically-generated hydrogen peroxide at 0.6 V versus Ag/AgCl. It showed a linear response ranging from 0.2 to 2.0 mM with a detection limit of 0.002 mM.

Recently, it was demonstrated that particles could be utilized as carrier systems for drugs into the hair follicles. In the present study, a two-component drug delivery system is presented consisting of degradable particles loaded with fluorescein isothiocyanate and a separate protease formulation for degradation. The particles were applied alone, 30 min previous to the protease application and simultaneously with the protease onto porcine skin. Subsequently, biopsies were removed, and the penetration depths of the particles were analyzed using laser scanning microscopy.

The obtained results demonstrate that the particles alone achieved a penetration depth of around 900 μm. Similar results were obtained for the successive application of particles and protease, whereas a release of the fluorescent dye was only observed in the upper 250 μm corresponding to the penetration depth of the protease. In the case of the simultaneous application, the particles were partly dissolved before application, leading to a reduced particle size and diminished penetration depth.

The results revealed that degradable particles are a promising tool for drug delivery into the skin.

The coordination of zinc by proteins and various other organic molecules is essential for numerous biological processes, such as in enzymatic catalysis, metabolism and signal transduction. Presence of small molecular chelants can have a profound effect on the bioavailability of zinc and affect critical Zn²⁺-protein interactions. Zn²⁺ chelators are also emerging therapeutics for Alzheimer’s diseases because of their preventive effect on zinc promoted amyloid formation. Despite the importance of zinc-protein-chelant interactions in biology and medicine, probing such interactions is  challenging. Here, we introduce an innovative approach for real-time characterization of zinc-protein-chelant interactions using gold nanoparticles (AuNPs) functionalized with a zinc-responsive protein mimetic polypeptide. The peptide functionalized AuNPs aggregate extensively in the presence of Zn²⁺, triggered by specific Zn²⁺-mediated polypeptide dimerization and folding, causing a massive red shift of the plasmon band. Chelants affects the Zn²⁺- polypeptide interaction and thus the aggregation differently depending on their concentrations, zincbinding affinities and coordination numbers, which affect the position of the plasmon band. This system is a simple and powerful tool that provides extensive information about the interactions of chelants in the formation of Zn²⁺ coordination complexes and is an interesting platform for development of bioanalytical techniques and characterization of chelation-based therapeutics.

A novel class of organic nanoparticles as biolabels that can generate an instant visible signal was applied to immunodipsticks. A new principle for signal generation based on hydrolysis of colourless signal precursor molecules to produce coloured signal molecules followed by signal precipitation and localization was demonstrated. The nanoparticle biolabels were applied to sandwich immunoassays for the detection of mouse immunoglobulin G (M IgG). In the presence of M IgG, a nanoparticle-immunocomplex was formed and bound on the test zone immobilized with goat anti M IgG (Gt α M IgG). A blue line was developed on the test zone upon the addition of a signal developing reagent. An optical signal could be simply assessed using naked eyes or quantified using a reading device. The lowest visible signal that could be observed using naked eyes was found to be 1.25 μg L⁻¹ M IgG. The nanoparticle biolabel also showed a better sensitivity (signal-to-noise ratio) compared with the conventional colloidal gold biolabel. This novel class of organic nanoparticles offers an alternative biolabel system for the development of point-of-care immunodipsticks.

We report on cell surface engineering of living microorganisms by using Layer-by-Layer (LbL) technology to extend the substrate spectrum. The yeast Arxula adeninivorans LS3 (Arxula) was employed as a model organism and biological template. By using LbL technology, Arxula cells were encapsulated by polyelectrolyte and enzyme layers. The biological activity of the Arxula was retained after the encapsulation process. The polymeric capsule surrounding the Arxula provides a stable interface for surface engineering of living cells. LbL of polyelectrolytes followed by an enzyme layer of lactate oxidase were assembled. The outer enzyme layer provides an additional biological function for Arxula to convert the unfavourable substrate lactate into the favourable substrate pyruvate, thus extending the substrate spectrum of the organism. Moreover, capsule stability and enzyme conjugate stability of the surface engineered Arxula were studied.

We report on using fluorescence microscopy to study, visualize and determine the diffusion phenomena into and bioaffinity binding within single microcapsules in real time by using biotin–fluorescein as diffusive species and encapsulated avidin as binding partner. Microcapsules were constructed by entrapment of avidin within an agarose matrix and encapsulated with polyelectrolyte layers by Layer-by-Layer (LbL) polyelectrolyte self assembly. A “ring” of high fluorescence intensity advancing with time towards the capsule centre was observed during incubation of capsules with fluorescent-labeled biotin. Fluorescence intensity was build up in capsule areas where binding to avidin occurred and was visualized in real time. A model for the diffusion process in microcapsules was developed and experimental data was plotted and fitted well with trends predicted by the model. The value of the diffusion coefficient for biotin–fluorescein was determined to be 3.5 × 10⁻⁸ cm²/s, which is comparable to literature values of similar sized molecules.

The tremendous growth of disposable electrode-based portable devices for point-of-care testing requires mass production of disposable electrodes in a low-cost and sustainable manner. Here, we demonstrate a green route for the conversion of biomass lignin, patterning, and reduction of the lignin-derived graphene electrodes by sequential laser lithography, water lift-off and sodium borohydride (NaBH4) treatment, and their use for electrochemical lactate biosensors. Energy-saving and localized laser lithography converted the aromatic ring-rich lignin into porous laser-induced graphene (LIG). The conductivity and attachment of the LIG to the substrate were optimized in a factorial experiment with laser power and scan speed as variables. Characterization results revealed the conversion of partial heteroatoms (e.g., Na, S, O) into granular inorganic compounds on the LIG surface under laser treatment. Water was used as an eco-friendly solvent for the patterning of the LIG (P-LIG) by a lift-off process, where the inorganic residues and un-reacted lignin were dissolved, exposing the macro-/micropores in the P-LIG. NaBH4 induced a reduction of the P-LIG (P-rLIG) resulting in improved electrochemical kinetics with lower charge transfer resistance (27.3 omega) compared to the LIG (248.1 omega) and the P-LIG (61.4 omega). The porous P-rLIG served as a 3D electrode for the deposition of Prussian blue and lactate oxidase for disposable electrochemical lactate biosensors, delivering a good analytical performance towards lactate detection with a linear range up to 16 mM and a high sensitivity (1.21 mu A mM-1). These lignin-derived disposable electrodes, utilizing renewable resources together with low-energy consumption fabrication and patterning, may contribute to the sustainable manufacturing of biosensors for point-of-care and point-of-use applications.