Current Trends of Nanobiosensors for Point-of-Care Diagnostics.

In order to provide better-quality health care, it is very important that high standards of health care management are achieved by making timely decisions based on rapid diagnostics, smart data analysis, and informatics analysis. Point-of-care testing ensures fast detection of analytes near to the patients facilitating a better disease diagnosis, monitoring, and management. It also enables quick medical decisions since the diseases can be diagnosed at an early stage which leads to improved health outcomes for the patients enabling them to start early treatment. In the recent past, various potential point-of-care devices have been developed and they are paving the way to next-generation point-of-care testing. Biosensors are very critical components of point-of-care devices since they are directly responsible for the bioanalytical performance of an essay. As such, they have been explored for their prospective point-of-care applications necessary for personalized health care management since they usually estimate the levels of biological markers or any chemical reaction by producing signals mainly associated with the concentration of an analyte and hence can detect disease causing markers such as body fluids. Their high selectivity and sensitivity have allowed for early diagnosis and management of targeted diseases; hence, facilitating timely therapy decisions and combination with nanotechnology can improve assessment of the disease onset and its progression and help to plan for treatment of many diseases. In this review, we explore how nanotechnology has been utilized in the development of nanosensors and the current trends of these nanosensors for point-of-care diagnosis of various diseases.

Green Synthesis and Characterization of Silver Nanoparticles Using Citrullus lanatus Fruit Rind Extract.

The wide-scale application of silver nanoparticles (AgNPs) in areas such as chemical sensing, nanomedicine, and electronics has led to their increased demand. Current methods of AgNPs synthesis involve the use of hazardous reagents and toxic solvents. There is a need for the development of new methods of synthesizing AgNPs that use environmentally safe reagents and solvents. This work reports a green method where silver nanoparticles (AgNPs) were synthesized using silver nitrate and the aqueous extract of Citrullus lanatus fruit rind as the reductant and the capping agent. The optimized conditions for the AgNPs synthesis were a temperature of 80°C, pH 10, 0.001 M AgNO3, 250 g/L watermelon rind extract (WMRE), and a reactant ratio of 4 : 5 (AgNO3 to WMRE). The AgNPs were characterized by Ultraviolet-Visible (UV-Vis) spectroscopy exhibiting a λmax at 404 nm which was consistent with the spectra of spherical AgNPs within the wavelength range of 380-450 nm, and Cyclic Voltammetry (CV) results showed a distinct oxidation peak at +291 mV while the standard reference AgNPs (20 nm diameter) oxidation peak occurred at +290 mV, and Transmission Electron Microscopy (TEM) revealed spherical shaped AgNPs. The AgNPs were found to have an average diameter of 17.96 ± 0.16 nm.

Biosensor for selective detection of E. coli in spinach using the strong affinity of derivatized mannose with fimbrial lectin.

Escherichia coli (E. coli) contamination in foods and water resources represents a major threat for human health and the environment. This work exploits the strong affinity of mannose-containing oligosaccharides with the fimbrial lectin of E. coli to design novel biosensors. Modified carbohydrate ligands were synthesized by introducing phenyl residues and aliphatic chains to mannose via reductive amination in order to increase both the affinity and selectivity to E. coli compared to other pathogenic bacteria. The synthesized ligands include p-thiolphenyl aminomannose (PTAM), p-carboxyphenyl aminomannose (PCAM), 1-deoxy-1-aminomannopyranoside (DAMP), glucosamine and low molecular weight chitosan bonded to mercapto undecanoic acid. The structures of the ligands were confirmed using (1)H NMR and 1H, (13)C, COZY NMR, and ESI/MS. The ligands were immobilized onto gold electrodes and SPR surfaces using-mercaptoundecanoic acid with glycine as deactivating agent. Two detection mechanisms were tested: (i) metal-enhanced electrochemical detection (MED) and (ii) label-free surface plasmon resonance (SPR) detection. The introduction of phenyl residues and aliphatic side groups to the mannose-containing oligosaccharides produced extremely high affinity for E. coli with detection limit of 1 cfu/mL. The relative selectivity of these ligands for E. coli, Citrobacter freundii, Staphylococcus epidermidis were 100%, 2.6% and 8.6% respectively. The biosensors were validated using spinach leaves at 3.0 cfu/mL. The work provides a generic biosensor for other pathogenic bacteria by enabling multivalent binding, immediate recognition for pathogens as well as inhibition of bacterial growth.

Conducting polyamic acid membranes for sensing and site-directed immobilization of proteins.

A biosensor platform based on polyamic acid (PAA) is reported for oriented immobilization of biomolecules. PAA, a functionalized conducting polymer substrate that provides electrochemical detection and control of biospecific binding, was used to covalently attach biomolecules, resulting in a significant improvement in the detection sensitivity. The biosensor sensing elements comprise a layer of PAA antibody (or antigen) composite self-assembled onto gold (Au) electrode via N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) linking. The modified PAA was characterized by Fourier transform infrared (FTIR), (1)H nuclear magnetic resonance (NMR), and electrochemical techniques. Cyclic voltammetry and impedance spectroscopy experiments conducted on electrodeposited PAA on Au electrode using ferricyanide produced a measurable decrease in the diffusion coefficient compared with the bare electrode, indicating some retardation of electron transfer within the bulk material of the PAA. Thereafter, the modified PAA surface was used to immobilize antibodies and then to detect inducible nitric oxide synthase and mouse immunoglobulin G (IgG) using enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), and amperometric techniques. ELISA results indicated a significant amplified signal by the modified PAA, whereas the SPR and amperometric biosensors produced significant responses as the concentration of the antigen was increased. Detection limits of 3.1×10(-3)ng/ml and 2.7×10(-1)ng/ml were obtained for SPR and amperometric biosensors, respectively.

Immunosensors for quantifying cyclooxygenase 2 pain biomarkers.

BACKGROUND: Cyclooxygenase 2 (COX-2) is a key enzyme in pain biomarkers, inflammation and cancer cell proliferation. Thus biosensors that can quantify pain mediators based on biochemical mechanism are imperative. METHODS: Biomolecular recognition and affinity of antigenic COX-2 with the antibody were investigated using surface plasmon resonance (SPR) and ultra-sensitive portable capillary (UPAC) fluorescence sensors. Polyclonal goat anti-COX-2 (human) antibodies were covalently immobilized on gold SPR surface and direct recognition for the COX-2 antigen assessed. The UPAC sensor utilized an indirect sandwich design involving covalently attached goat anti-COX-2 as the capture antibody and rabbit anti-COX-2 (human) antibody as the secondary antibody. RESULTS: UPAC fluorescence signals were directly proportional to COX-2 at a linear range of 7.46×10⁻4-7.46×10¹ ng/ml with detection limit of 1.02×10⁻4 ng/ml. With SPR a linear range was 3.64×10⁻4-3.64×10² ng/ml was recorded and a detection limit of 1.35×10⁻4 ng/ml. Validation was achieved in simulated blood samples with percent recoveries of 81.39% and 87.23% for SPR and UPAC respectively. CONCLUSION: The developed sensors have the potential to provide objective characterization of pain biomarkers for clinical diagnoses.

Detection of inducible nitric oxide synthase using a suite of electrochemical, fluorescence, and surface plasmon resonance biosensors.

A suite of biosensors for rapid detection of inducible nitric oxide synthase (iNOS) is described. First, a metal-enhanced electrochemical detection (MED) sensor, which relied on the redox properties of a silver monolayer, was developed. The linear detection range was between 8.64×10(-2) and 5.4×10(1)ng/ml with a detection limit of 1.69×10(-4)ng/ml. This method was compared with surface plasmon resonance (SPR) biosensors in which polyclonal mouse anti-iNOS was covalently immobilized onto a gold surface using an iNOS antigen. The linear detection range recorded was between 3.37×10(1) and 5.4×10(-2)ng/ml with a detection limit of 2×10(-3)ng/ml. Finally, an ultrasensitive portable capillary (UPAC) fluorescence immunosensor, in which a mouse anti-iNOS antibody was covalently immobilized onto the inner surface of a capillary and a rabbit anti-iNOS antibody was employed as the secondary antibody, was developed. The resulting signals were found to be directly proportional to iNOS concentrations between 1.52×10(-1) and 1.52×10(-2)ng/ml with a detection limit of 1.05×10(-3)ng/ml. These immunosensors exhibit low cross-reactivity toward potential interferents such as human serum albumin and ovalbumin. The SPR and UPAC biosensors were validated using simulated blood spiked with recombinant iNOS, resulting in recoveries of 85% and 88.5%, respectively. The research presented in this article could potentially provide new ways of detecting NO for diagnostic and biomarker purposes in medical research.