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An optical biosensor for label-free detection of ochratoxin A (OTA) in beer samples is presented. The biosensor consists of an array of ten Mach-Zehnder interferometers (MZIs) monolithically integrated along with their respective broad-band silicon light sources on the same Si chip (37 mm2). The chip was transformed to biosensor by functionalizing the MZIs sensing arms with an OTA-ovalbumin conjugate. OTA determination was performed by pumping over the chip mixtures of calibrators or samples with anti-OTA antibody following a competitive immunoassay format. An external miniaturized spectrometer was employed to continuously record the transmission spectra of each interferometer. Spectral shifts obtained due to immunoreaction were transformed to phase shifts through Discrete Fourier Transform. The assay had a detection limit of 2.0 ng/ml and a dynamic range 4.0–100 ng/ml in beer samples, recoveries ranging from 90.6 to 116%, and intra- and inter-assay coefficients of variation of 9% and 14%, respectively. The results obtained with the sensor using OTA-spiked beer samples spiked were in good agreement with those obtained by an ELISA developed using the same antibody. The good analytical performance of the biosensor and the small size of the proposed chip provide for the development of a portable instrument for point-of-need determinations. © 2016 Elsevier B.V.

The label-free detection of bovine milk in goat milk through a miniaturized optical biosensor is presented. The biosensor consists of ten planar silicon nitride wave-guide Broad-Band Mach–Zehnder interferometers (BB-MZIs) monolithically integrated and self-aligned with their respective silicon LEDs on the same Si chip. The BB-MZIs were transformed to biosensing transducers by functionalizing their sensing arm with bovine k-casein. Measurements were performed by continuously recording the transmission spectra of each interferometer through an external spectrometer. The amount of bovine milk in goat milk was determined through a competitive immunoassay by passing over the sensor mixtures of anti-k-casein antibodies with the calibrators or the samples. The output spectra of each BB-MZI recorded during the reaction were subjected to Discrete Fourier Transform in order to convert the observed spectral shifts to phase shifts in the wave-number domain. The method had a detection limit of 0.04 % (v/v) bovine milk in goat milk, dynamic range 0.1–1.0 % (v/v), recoveries 93–110 %, and intra-and inter-assay coefficients of variation less than 12 and 15 %, respectively. The proposed biosensor compared well in terms of analytical performance with a competitive ELISA developed using the same monoclonal antibodies. Nevertheless, the duration of the biosensor assay was 10 min whereas the ELISA required 2 h. Thus, the fast and sensitive determinations along with the small size of the sensor make it ideal for incorporation into portable devices for assessment of goat or ewe’s milk adulteration with bovine milk at the point-of-need. © Springer-Verlag Berlin Heidelberg 2015.

The early diagnosis of acute myocardial infarction requires the determination of several markers in serum shortly after its incidence. The markers most widely employed are the isoenzyme MB of creatine kinase (CK-MB) and the cardiac troponin I (cTnI). In the present work, a capillary waveguide fluoroimmunosensor for fast and highly sensitive simultaneous determination of these markers in serum samples is demonstrated. The dual-analyte immunosensor was realized using glass capillaries internally modified with an ultrathin poly(dimethylsiloxane) film by creating discrete bands of analyte-specific antibodies. The capillary was then filled with a mixture of sample and biotinylated detection antibodies followed by reaction with streptavidin- horseradish peroxidase and incubation with a fluorescently labeled tyramide derivative to accumulate fluorescent labels onto immunoreaction bands. Upon scanning the capillary with a laser beam, part of the emitted fluorescence is trapped and waveguided through the capillary wall to a photomultiplier placed on one of its ends. The employment of tyramide signal amplification provided detection limits of 0.2 and 0.5 ng/mL for cTnI and CK-MB, respectively, in a total assay time of 30 min compared to 0.8 and 0.6 ng/mL obtained for the corresponding assays when the conventional fluorescent label R-phycoerythrin was used in a 65-min assay. In addition, the proposed immunosensor provided accurate and repeatable measurements (intra-assay and interassay coefficients of variation lower than 10%), and the values determined in serum samples were in good agreement with those obtained with commercially available enzyme immunoassays. Thus, the proposed capillary waveguide fluoroimmunosensor has all the required characteristics for fast and reliable diagnosis of acute myocardial infarction. © 2009 Springer-Verlag.

An optical capillary waveguide fluoroimmunosensor based on glass capillaries internally coated with an ultrathin poly(dimethylsiloxane) (PDMS) film is presented. The evaluation of the capillaries developed was done in comparison with aminosilanized [3-(aminopropyl)triethoxysilane, APTES] glass and poly(methylpentene) (PMP) capillaries by immobilizing rabbit γ-globulins on the internal capillary wall. Following reaction with (R)-phycoerythrin- labelled antibody, the capillary was scanned with a laser beam and the fluorescence waveguided through the capillary wall was detected by a photomultiplier placed at one of its ends. The capillaries developed provided considerably improved protein coating homogeneity (intracapillary coefficients of variation 2.9-6.6%) and repeatability (intercapillary coefficients of variation 2.1-5.0%) compared with APTES-treated ones (7.9-13.4 and 8.5-15.2%, respectively). With use of these capillaries in a sandwich-type immunosensor for the determination of rabbit γ-globulins, the assay detection limit was improved eightfold (4.4 ng/mL) compared with that obtained using PMP capillaries (35.3 ng/mL), whereas the assay repeatability was improved threefold (intra-assay coefficients of variation 5.9-13.1%) compared with APTES-treated capillaries (15.6-36%). [Figure not available: see fulltext.] © 2008 Springer-Verlag.

Methods for homogeneous coverage of surfaces with biomolecules are critical for the development of bioanalytical microsystems. Here we present a simple method for coating glass capillaries with an ultra-thin poly(dimethylsiloxane) (PDMS) film in order to be used as optical waveguide fluoroimmunosensors. A model rabbit γ-globulin binding assay was employed for the evaluation of modified capillaries in terms of protein binding capacity and protein coating homogeneity. It was found that the PDMS-modified capillaries provided excellent protein coating homogeneity inside each one capillary (CVs 2.9-6.6%) and between-capillaries repeatability (CVs 2.1-5.0%) compared to aminosilanized ones. Furthermore, the waveguiding properties of the glass were not affected by the proposed surface modification. © 2008 Elsevier B.V. All rights reserved.

Journal URL: http://springerlink.metapress.com/content/100417

An optical real-time affinity biosensor, which is based on a monolithic silicon optoelectronic transducer and a microfluidic module, is described. The transducer monolithically integrates silicon avalanche diodes as light sources, silicon nitride optical fibers, and p/n junction detectors and efficiently intercouples these elements through a self-alignment technique. The transducer surface is hydrophilized by oxygen plasma treatment, silanized with (3-aminopropyl)triethoxysilane and bioactivated through adsorption of the biomolecular probes. The use of a microfluidic module allows real-time monitoring of the binding reaction of the gold nanoparticle-labeled analytes with the immobilized probes. Their binding within the evanescent field at the surface of the optical fiber causes attenuated total reflection of the waveguided modes and reduction of the detector photocurrent. The biotin-streptavidin model assay was used for the evaluation of the analytical potentials of the device developed. Detection limits of 3.8 and 13 pM in terms of gold nanoparticle-labeled streptavidin were achieved for continuous- and stopped-flow assay modes, respectively. The detection sensitivity was improved by silver plating of the immolilized gold nanoparticles, and a detection limit of 20 fM was obtained after 20-min of silver plating. In addition, two different analytes, streptavidin and anti-mouse IgG, were simultaneously assayed on the same chip demonstrating the multianalyte potential of the sensor developed.

The development of a four-band capillary optical immunosensor for the simultaneous determination of mesotrione, hexaconazole, paraquat, and diquat is described. Four distinct bands (each corresponding to a different analyte) are created in the internal walls of a plastic capillary by immobilizing protein conjugates of the analytes. To perform the assay, the capillary is filled with a mixture of antianalyte-specific antibodies together with a standard or sample containing the analyte(s). After a short incubation, a mixture of the appropriate second antibodies labeled with fluorescein is introduced into the capillary. To measure the fluorescence intensity bound onto each band, the capillary was scanned, perpendicularly to its axis, by a laser light beam. Part of the emitted photons were trapped into the capillary walls and waveguided to a photomultiplier placed at the one end of the capillary. The analytical characteristics of the assays of mesotrione, paraquat, diquat, and hexaconazole were as follows: detection limits of 0.04, 0.06, 0.09, and 0.10 ng/mL, respectively; dynamic ranges up to 9, 6, 12, and 15 ng/ mL, respectively; intra- and interassay CVs less than 10%. The analytical characteristics of the assays were comparable with those of the corresponding single-analyte fluoroimmunoassays performed in microtitration wells, proving the ability of the proposed immunosensor for reliable multianalyte determinations. Moreover, the combination of low-cost disposable plastic capillary tubes with the low consumption of reagents, the short assay time, and the multianalyte feature of the proposed immunosensor indicates its potential for environmental analysis.