The use of gold nanoparticles (AuNPs) as labeling carriers in combination with the enzymatic activity of the Horseradish Peroxidase (HRP) in order to achieve an improved optical lateral flow immunoassay (LFIA) performance is here presented.Briefly in a LFIA with an immune-sandwich format AuNPs are functionalized with a detection antibody already modified with HRP, obtaining an "enhanced" label. Two different detection strategies have been tested: the first one following just the red color of the AuNPs and the second one using a substrate for the HRP (3 different substrates are evaluated), which produces a darker color that enhances the intensity of the previous red color of the unmodified AuNPs. In such very simple way it is gained sensitivity (up 3

Selective transport in nanochannels (protein-based ion channels) is already used in living systems for electrical signaling in nerves and muscles, and this natural behavior is being approached for the application of biomimetic nanochannels in biosensors. On the basis of this principle, single nanochannels and nanochannel arrays seem to bring new advantages for biosensor development and applications. The purpose of this review is to provide a general comprehensive and critical overview on the latest trends in the development of nanochannel-based biosensing systems. A detailed description and discussion of representative and recent works covering the main nanochannel fabrication techniques, nanoporous material characterizations, and especially their application in both electrochemical and optical sensing systems is given. The state-of-the-art of the developed technology may open the way to new advances in the integration of nanochannels with (bio)molecules and synthetic receptors for the development of novel biodetection systems that can be extended to many other applications with interest for clinical analysis, safety, and security as well as environmental and other industrial studies and applications.

A rapid nanochannel-based immunoassay capable of the filtering and subsequent detection of proteins in whole blood without any sample preparation is described. This is accomplished by using a nanoporous/nanochannel membrane modified with antibodies, the conductivity of which toward a redox indicator is tuned by primary and secondary immunoreactions with proteins and gold nanoparticles. This interesting nanopore blockage by gold nanoparticles is enhanced by silver deposition that further decreases the diffusion of the signaling indicator through the nanochannel. The efficiency of the nanochannels to act as immunoreaction platforms including the use of nanoparticles is also monitored by microscopic techniques. Successful detection of immunoglobulins including a cancer biomarker is achieved in buffer as well as in whole blood. This system constitutes an efficient immunoassay capable of detecting up to 52 U mL(-1) of CA15-3. The developed nanochannel/nanoparticle-based device can be used for several other proteins and extended also to DNA detection with interest not only for diagnostics but also environmental monitoring, food analysis, safety, and security applications.

Although lateral flow assays (LFA) are currently being used in some point-of-care applications (POC) they cannot still be extended to a broader range of analytes for which higher sensitivities and lower detection limits are required. To overcome such drawbacks, we propose here a simple and facile alternative based on the use of delay hydrophobic barriers fabricated by wax-printing so as to improve the LFA sensitivity. Several wax pillars patterns were printed onto nitrocellulose membrane in order to produce delays as well as pseudo turbulences into the microcapillary flow. The effect of the proposed wax pillar modified devices were also mathematically simulated corroborating the experimental results obtained for the different patterns tested afterwards for detection of HIgG as model protein in a gold nanoparticle-based LFA. The effect of the introduction of such wax-printed pillars was the sensitivity improvement of almost 3-folds in comparison to a conventional free-barrier LFA.

Infectious plant diseases are caused by pathogenic microorganisms such as fungi, bacteria, viruses, viroids, phytoplasma and nematodes. Worldwide, plant pathogen infections are among main factors limiting crop productivity and increasing economic losses. Plant pathogen detection is important as first step to manage a plant disease in greenhouses, field conditions and at the country boarders. Current immunological techniques used to detect pathogens in plant include enzyme-linked immunosorbent assays (ELISA) and direct tissue blot immunoassays (DTBIA). DNA-based techniques such as polymerase chain reaction (PCR), real time PCR (RT-PCR) and dot blot hybridization have also been proposed for pathogen identification and detection. However these methodologies are time-consuming and require complex instruments, being not suitable for in-situ analysis. Consequently, there is strong interest for developing new biosensing systems for early detection of plant diseases with high sensitivity and specificity at the point-of-care. In this context, we revise here the recent advancement in the development of advantageous biosensing systems for plant pathogen detection based on both antibody and DNA receptors. The use of different nanomaterials such as nanochannels and metallic nanoparticles for the development of innovative and sensitive biosensing systems for the detection of pathogens (i.e. bacteria and viruses) at the point-of-care is also shown. Plastic and paper-based platforms have been used for this purpose, offering cheap and easy-to-use really integrated sensing systems for rapid on-site detection. Beside devices developed at research and development level a brief revision of commercially available kits is also included in this review.

Herein, we describe the development of a new, highly efficient label for immunosensing comprising an antibody/PEGylated gold-nanoparticle (AuNP) conjugate in which the antibody molecules are bound to the AuNP surface in a fixed orientation. Our method exploits the high density of positive charges on the major plane of antibodies that exists when the pH of the solution is lower than the isoelectric point of the antibody; the antibody molecules interact with the negatively charged AuNP surface through their major plane, enabling the antigen binding sites to move freely and therefore to reach maximum accessibility. This directed ionic interaction is reinforced by the formation of a peptide bond between the amino group of the Lys residues in the antibodies and the carboxylic groups of the PEGylated-AuNP surface via EDC chemistry. Electrochemical analyses revealed that the fixed-orientation conjugate offers a limit of detection that is 1 order of magnitude lower than that of a randomly oriented label. The performance of the new conjugate as an immunosensing label was assessed for the quantitative detection of IgG in human serum.

Lateral flow immunoassays (LFIA) are ideal biosensors to detect proteins, but their lack of sensitivity hinders their extensive use. We report a strategy that yields up to an 8-fold improvement in the sensitivity of a gold nanoparticles-based LFIA by changing the sizes of the pads. Theoretical flow simulations of the developed LFIA architectures are in accordance with the experimental results.

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