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PubMed2002

Whole-cell biosensing of 3-chlorocatechol in liquids and soils.

A rapid and sensitive technique is needed to analyze water and soils for chlorocatechols, common environmental pollutants produced from wood pulp chlorination and other processes. The soil bacteria Pseudomonas putida, harboring plasmid pSMM50R-B', selectively express beta-galactosidase in response to 3-chlorocatechol in pure water samples. The objective of the study was to determine whether background matrices in fresh water, sea water, soils, and organic solvents interfered with 3-chlorocatechol analysis by use of a bacteria-sensing system and by high-performance liquid chromatography (HPLC). Although 3-chlorocatechol detection by HPLC was not substantially affected by the background composition of aqueous or organic solvents, HPLC was ineffective in the analysis of contaminated soils due to irreversible contaminant sorption. Whereas detection by the bacteria-sensing system was reduced in the presence of aqueous and organic solvents, interferences could be reduced by sample dilution. 3-Chlorocatechol was detected when the bacteria were added directly to contaminated soils, suggesting that the organism enhanced desorption or had access to the sorbed compounds. Results indicate that the bacteria-sensing system has wide application for detection of 3-chlorocatechols in environmental samples, especially in soils where extraction and HPLC analysis are not efficient due to extensive contaminant sorption.

Catechols↗
PubMed2006

Multi-analyte SPR immunoassays for environmental biosensing of pesticides.

Multi-analyte detection of environmentally relevant pesticides is performed by using a two-channelled surface plasmon resonance (SPR) biosensor. The special design of the SPR instrument allows the determination of several analytes (DDT, chlorpyrifos and carbaryl) via different immobilization formats. First, simultaneous pesticide monitoring is possible by flowing chlorpyrifos, carbaryl or DDT samples separately over each channel of the SPR system, wherein their corresponding recognition element was previously immobilized. The second approach is based on the multiple and combined immobilization of several analyte recognition elements on the sensing surface of one individual flow cell. In this format, the analysis time for all three pesticides varied from 40 to 60 min depending on the number of regeneration cycles. In most cases, similar detection limits were attained for the target analyte irrespective of the assay format, with sensitivity values at the nanogram per litre level (18-50 ng L(-1)). The assay reproducibility was proved through the repeated use of the same sensor surface for over more than 200 assay cycles, whereas the absence of biosensor response to non-related analytes showed the specificity and reliability of the analysis. The SPR instrument, including optics, electronics and microfluidics, is already commercialised by the company SENSIA, SL.

Antibodies↗
PubMed2005

Sensitivity enhancement of surface plasmon resonance biosensing of small molecules.

Surface plasmon resonance (SPR) biosensor formats using gold nanoparticle or protein signal amplification for the sensitive assay of small molecules were developed using progesterone as a model compound. Progesterone was immobilized to a dextran surface in the Biacore biosensor through in situ covalent immobilization using an oligoethylene glycol linker attached to the 4 position of the steroid. This surface produced stable antibody binding for in excess of 1100 assay cycles. Using this surface, assays were developed for progesterone using 10- and 20-nm gold-streptavidin labels attached to biotinylated monoclonal antibody in both label prebinding and sequential binding formats. Prelabeling formats gave no signal enhancement but produced assays with limits of detection of 143 pg/ml, compared with approximately 1 ng/ml in previous studies. Sequential binding formats gave signal enhancements of 2.2-fold over the monoclonal antibody and a limit of detection of 23.1 pg/ml. It was found that secondary antibody labeling gave 8.1-fold signal enhancements and a limit of detection of 20.1 pg/ml, whereas use of secondary antibody-25 nm gold complexes provided more signal enhancement (13-fold) and a further improvement in limit of detection of 8.6 pg/ml.

Antibodies, Monoclonal↗
PubMed2013

WITHDRAWN: Interferometric biosensing of DNA-damaging chemicals.

This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

Journal Article↗
PubMed2006

Estrogen conjugation and antibody binding interactions in surface plasmon resonance biosensing.

Thioether-linked 3-mercaptopropionic acid derivatives of 17beta-estradiol and estrone were formed at the A-ring 4-position of the steroids by substitution of their 4-bromo analogues. The carboxylic acid terminal was used to link to an oligoethylene glycol (OEG) chain of 15-atoms in length. The OEG derivative of 17beta-estradiol was then in situ immobilized on a carboxymethylated dextran-coated gold sensor surface used to detect refractive index changes upon protein binding to the surface by surface plasmon propagation in a BIAcore surface plasmon resonance (SPR) instrument. Two other estradiol-OEG derivatives with Mannich reaction linkage at the 2-position and hemisuccinate linkage at the 3-position were also immobilized on the sensor surfaces for comparison. Binding performance between these immobilized different positional conjugates and monoclonal anti-estradiol antibody, raised from a 6-position conjugate, clearly demonstrated that both 2- and 4-conjugates, not conjugated through existing functional groups, gave strong antibody bindings, whereas the 3-conjugate through an existing functional group (3-OH) gave very little binding (2% compared to the 2-conjugate). Both 2- and 4-position conjugates were then applied in a highly sensitive estradiol SPR immunoassay with secondary antibody mediated signal enhancement that gave up to a 9.5-fold signal enhancement of primary antibody binding, and a detection limit of 25 pg/mL was achieved for a rapid and convenient flow-through immunoassay of estradiol.

Antibodies↗
PubMed2004

Label-free biosensing by surface plasmon resonance of nanoparticles on glass: optimization of nanoparticle size.

The unique optical properties of noble metal nanoparticles have been used to design a label-free biosensor in a chip format. In this paper, we demonstrate that the size of gold nanoparticles significantly affects the sensitivity of the biosensor. Gold nanoparticles with diameters in the range of 12-48 nm were synthesized in solution and sensor chips were fabricated by chemisorption of these nanoparticles on amine-functionalized glass. Sensors fabricated from 39-nm-diameter gold nanoparticles exhibited maximum sensitivity to the change of the bulk refractive index and the largest "analytical volume", defined as the region around the nanoparticle within which a change in refractive index causes a change in the optical properties of the immobilized nanoparticles. The detection limit for streptavidin-biotin binding of a sensor fabricated from 39-nm-diameter nanoparticles was 20-fold better than a previously reported sensor fabricated from 13-nm-diameter gold nanoparticles. We also discuss several other factors that could improve the performance of the next generation of these immobilized metal nanoparticle sensors.

Glass↗
PubMed2004

Spongiform immobilization architecture of ionotropy polymer hydrogel coentrapping alcohol oxidase and horseradish peroxidase with octadecylsilica for optical biosensing alcohol in organic solvent.

An organic-phase optical alcohol biosensor consisting of alcohol oxidase and horseradish peroxidase coimmobilized in a spongiform hydrogel matrix of hydroxethyl carboxymethyl cellulose, an adduct of 3-methoxy-4-ethoxy benzaldehyde, 4-tert-butylpyridinium acetohydrazone, silica gel particles, and octadecylsilica particles in conjunction with an optical oxygen transducer has been successfully fabricated. The novel enzyme entrapment structure was mainly characterized with desirable solvent permeability, high efficiency of mass transfer for reactants, and good accessibility and stability of the immobilized enzymes. The biosensor could work in water-miscible solvent such as a solvent mixture of acetonitrile and phosphate aqueous buffer, as well as hydrophobic organic solvent such as n-hexane. The biosensor had the highest sensitivity to methanol in both solvent systems. Under the stop-flow mode, the biosensor had the analytical working ranges from 80 microM to 90 mM methanol in n-hexane and 0.10 to 90 mM methanol in acetonitrile/buffer. When the biosensor functioned in n-hexane, it could take benzaldehyde as an alcohol substrate and was free from any pH disturbance. In the presence of coimmobilized horseradish peroxidase, the operational life of the biosensor was 60 assays and the shelf life was longer than two weeks. The biosensor has been satisfactorily applied to the determination of methanol in commercial gasoline-methanol blend samples.

Alcohol Oxidoreductases↗
PubMed2001

Pulse mode operation of Love wave devices for biosensing applications.

In this work we present a novel pulse mode Love wave biosensor that monitors both changes in amplitude and phase. A series of concentrations of 3350 molecular weight poly(ethylene glycol) (PEG) solutions are used as a calibration sequence for the pulse mode system using a network analyzer and high frequency oscilloscope. The operation of the pulse mode system is then compared to the continuous wave network analyzer by showing a sequence of deposition and removal of a model mass layer of palmitoyl-oleoyl-sn-glycerophosphocholine (POPC) vesicles. This experimental apparatus has the potential for making many hundreds of measurements a minute and so allowing the dynamics of fast interactions to be observed.

Journal Article↗
PubMed2002

Biosensing of arteriosclerotic nanoplaque formation and interaction with an HMG-CoA reductase inhibitor.

Proteoheparan sulphate can be adsorbed to a methylated silica surface in a monomolecular layer via its transmembrane hydrophobic protein core domain. As a result of electrostatic repulsion, its anionic glycosaminoglycan side chains are stretched out into the blood substitute solution, thereby representing one receptor site for specific lipoprotein binding through basic amino acid-rich residues within their apolipoproteins. The binding process was studied by ellipsometric techniques suggesting that high-density lipoprotein (HDL) has a high binding affinity and a protective effect on interfacial heparan sulphate proteoglycan layers with respect to low-density lipoprotein (LDL) and Ca2+ complexation. Low-density lipoprotein was found to deposit strongly at the proteoheparan sulphate-coated surface, particularly in the presence of Ca2+, apparently through complex formation 'proteoglycan-LDL-calcium'. This ternary complex build-up may be interpreted as arteriosclerotic nanoplaque formation on the molecular level responsible for the arteriosclerotic primary lesion. On the other hand, HDL bound to heparan sulphate proteoglycan protected against LDL deposition and completely suppressed calcification of the proteoglycan-lipoprotein complex. In addition, HDL was able to decelerate the ternary complex deposition. Therefore, HDL attached to its proteoglycan receptor sites is thought to raise a multidomain barrier, selection and control motif for transmembrane and paracellular lipoprotein uptake into the arterial wall. Although much remains unclear regarding the mechanism of lipoprotein depositions at proteoglycan-coated surfaces, it seems clear that the use of such systems offers possibilities for investigating lipoprotein deposition at a 'nanoscopic' level under close to physiological conditions. In particular, Ca2+-promoted LDL deposition and the protective effect of HDL even at high Ca2+ and LDL concentrations agree well with previous clinical observations regarding risk and beneficial factors for early stages of atherosclerosis. Considering this, the system was tested on its reliability in a biosensor application in order to unveil possible acute pleiotropic effects of the lipid lowering drug fluvastatin. The very low-density lipoprotein (VLDL)/intermediate-density lipoprotein (IDL)/LDL plasma fraction from a high risk patient with dyslipoproteinaemia and type 2 diabetes mellitus showed beginning arteriosclerotic nanoplaque formation already at a normal blood Ca2+ concentration, with a strong increase at higher Ca2+ concentrations. Fluvastatin, whether applied to the patient (one single 80 mg slow release matrix tablet) or acutely in the experiment (2.2 micromol L-1), markedly slowed down this process of ternary aggregational nanoplaque complexation at all Ca2+ concentrations used. This action resulted without any significant change in lipid concentrations of the patient. Furthermore, after ternary complex build-up, fluvastatin, similar to HDL, was able to reduce nanoplaque adsorption and size. These immediate effects of fluvastatin have to be taken into consideration while interpreting the clinical outcome of long-term studies.

Adsorption↗
PubMed2003

Planar silicon nitride waveguides for biosensing.

The principles of attenuation of the light intensity due to multiple reflections are realised in a planar silicon oxide (SiO(2))silicon nitride (Si(3)N(4)) waveguiding structure for the purpose of developing optical biosensors with improved sensitivity. The analysis of the experimental data shows that the large difference in refractive indices of core and cladding layers gives rise to an increase in sensitivity by a factor of 3 over previously reported structures. Composite polyelectrolyte self-assembled thin films containing cyclo-tetra-chromotropylene as an indicator and enzymes glucose oxidase or urease were employed in the superstrate as a sensing membrane. Individual enzyme reactions as well as their inhibition by pesticides were studied by monitoring the intensity of light output from the planar waveguide. The results were compatible with those obtained by conventional ultraviolet-visible absorption spectroscopy. The instrument detection limit for Imidacloprid pesticide was found to be as low as 10 ppb in concentration.

Journal Article↗
PubMed1995

Optical probes and transducers.

Biosensors are by definition a combination of a biological receptor compound and a physical or physicochemical transducer. Therefore, the transducing structure is a critical part of every biosensor. In the development of new and improved biosensing layers the importance of the transducing structure is not restricted to the substrate to which biological structures have to be coupled. A field of even greater importance is the use of transducers as probes providing information on the structure and function of biosensing layers, and their relation to a transducer surface. The aim of this paper is to give an overview on optical transducer principles and optical (surface) analytical techniques relevant as part of biosensing structures as well as probes in the development and optimisation of biosensing layers. Categories discussed are basic optical effects, materials involved, surface chemistry, the principal and technological limits of spatial resolution, and sensitivity. The intimate relation between the spatial resolution of a probe, the resulting size of interaction areas, and the feasibility of array structures is pointed out. Two interferometric methods are presented in principle, and their application to biosensing and some results are discussed in detail. The necessity to characterise receptor layers to get detailed information about the interaction process is pointed out. The close relationship between optimal characterisation of layers by selection of adequate probe technologies and improvement of probe performance, and the development of new biosensing layers is discussed. Finally, an outlook is given for future aspects of improved spatial resolution and multianalyte detection.

Biosensing Techniques↗
PubMed2004

Whole-cell-based biosensors for environmental biomonitoring and application.

A variety of whole-cell-based biosensors has been developed using numerous native and recombinant biosensing cells. The use of reporter genes, for example bacterial luciferase and gfp, to monitor gene expression is discussed in terms of each reporters' benefits and disadvantages, including their possible use on-line, their sensitivity, the need for extra substrate, etc. All biosensing cells in use can be classified into two groups in terms of their biosensing mechanisms--constitutive expression and stress- or chemical-specific inducible expression. In this review several examples of each are presented and discussed. The use of recombinant whole-cell biosensors in the field requires three components--biosensing cells, a measurement device, and a signal-transducing apparatus, the last two depending on the first and the final applications of the system. The use of different immobilization techniques in several studies to maintain the cells and their viability is also discussed, in particular their use in the development of both high-throughput and chip-based biosensing systems. Finally the application of whole-cell-based biosensors to different environmental media, such as water, soil, and atmospheric monitoring is discussed; particular attention is given to their use for detection of various stressors, including dioxins, endocrine-disrupting chemicals, and ionizing radiation.

Biosensing Techniques↗
PubMed2003

Evaluation of myocardial perfusion and left ventricular function six months after percutaneous transmyocardial laser revascularization: comparison of two Ho-YAG laser systems with the same wavelength, but different energy delivery and navigation systems.

BACKGROUND AND OBJECTIVES: Myocardial perfusion and left ventricular function (LVF) were assessed after percutaneous transmyocardial laser revascularization (PTMR) in patients not amenable to conventional revascularization, with a comparison of two laser systems. STUDY DESIGN/MATERIALS AND METHODS: PTMR was performed with an Eclipse laser in 15 patients, and with a Biosense DMR in 10 patients. (201)Thallium scintigraphy, coronary angiography, and ventriculography were performed at baseline and at the 7.5+/-4.3-month follow-up. All patients in the Biosense DMR group and 10 in the Eclipse group underwent NOGA mapping before PTMR and after follow-up. RESULTS: The event-free survival rates were comparable, and the angina scores of all patients improved significantly, but more so in the Biosense DMR group than in the Eclipse group (1.2+/-1.1 vs. 2.3+/-0.9, P < 0.05). Both, the electrical activity assessed by NOGA mapping and the normalized (201)thallium uptake at redistribution improved significantly in the treated segments after Biosense DMR, while the global LVF decreased insignificantly in the Eclipse group. CONCLUSIONS: PTMR resulted in significant improvements in the clinical symptoms, but the electrical activity improved only in the Biosense DMR group, without transforming to a better LVF.

Aged↗
PubMed2000

A liquid crystal pixel array for signal discrimination in array biosensors.

A new optical design uses a liquid crystal pixel array (LCPA) to discriminate multiple fluorescence signals on a two-dimensional biosensor array. The LCPA can selectively control the transmission of fluorescence generated from multiple biosensing elements on a planar waveguide. This device sequentially acquires the fluorescence data from the substrate by making multiple individual measurements of the sensing elements on the waveguide. The biosensing elements are patterned according to the pixel layout of the LCPA and optically aligned so that each electronically driven pixel can either transmit or filter out the fluorescence signal as specified by the user. The primary advantage of this system is that a single detection channel (i.e. photomultiplier tube (PMT)) can be used to measure multiple fluorescence signals from a two-dimensional substrate while the LCPA provides for spatial resolution. We evaluate the performance of the LCPA by testing the optical homogeneity of the liquid crystal pixels and linear dynamic range for transmitting light. The LCPA is also used with well-developed biosensing chemistry modified for this optical format.

Biosensing Techniques↗