scholarly journals Microarrays as Model Biosensor Platforms to Investigate the Structure and Affinity of Aptamers

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Jennifer A. Martin ◽  
Yaroslav Chushak ◽  
Jorge L. Chávez ◽  
Joshua A. Hagen ◽  
Nancy Kelley-Loughnane
Keyword(s):  
Minimum Distance ◽  
Limit Of Detection ◽  
Signal To Noise Ratio ◽  
Protein Labeling ◽  
Target Interaction ◽  
Starting Conditions ◽  
Recognition Elements ◽  
Labeling Method ◽  
Biosensor Applications ◽  
Magnitude Improvement

Immobilization of nucleic acid aptamer recognition elements selected free in solution onto the surface of biosensor platforms has proven challenging. This study investigated the binding of multiple aptamer/target pairs immobilized on a commercially available microarray as a model system mimicking biosensor applications. The results indicate a minimum distance (linker length) from the surface and thymine nucleobase linker provides reproducible binding across varying conditions. An indirect labeling method, where the target was labeled with a biotin followed by a brief Cy3-streptavidin incubation, provided a higher signal-to-noise ratio and over two orders of magnitude improvement in limit of detection, compared to direct Cy3-protein labeling. We also showed that the affinities of the aptamer/target interaction can change between direct and indirect labeling and conditions to optimize for the highest fluorescence intensity will increase the sensitivity of the assay but will not change the overall affinity. Additionally, some sequences which did not initially bind demonstrated binding when conditions were optimized. These results, in combination with studies demonstrating enhanced binding in nonselection buffers, provided insights into the structure and affinity of aptamers critical for biosensor applications and allowed for generalizations in starting conditions for researchers wishing to investigate aptamers on a microarray surface.

Development of a solid-phase extraction – spectrofluorimetric method for trace glutathione determination

2011 ◽  
Vol 89 (4) ◽  
pp. 517-523 ◽  
Author(s):  
Ke-Jing Huang ◽  
Cong-Hui Han ◽  
Ying-Ying Wu ◽  
Chao-Qun Han ◽  
De-Jun Niu ◽  
...  
Keyword(s):  
Solid Phase Extraction ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Signal To Noise Ratio ◽  
Calibration Graph ◽  
Extraction Column ◽  
Phase Extraction ◽  
Selective Reaction ◽  
Spectrofluorimetric Method ◽  
Relative Standard

A simple and efficient solid-phase extraction – spectrofluorimetric method has been developed to determine glutathione (GSH). Fluorescent probe N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl)iodoacetamide (BODIPY Fl-C1-IA) was used as the derivatization reagent. The procedure was based on a BODIPY Fl-C1-IA selective reaction with GSH to form the highly fluorescent product BODIPY Fl-C1-IA–GSH, using a solid-phase extraction column and spectrofluorimetric determination. The variables affecting analytical performance were studied and optimized. The calibration graph using the preconcentration system for GSH was linear over the range of 1–200 nmol/L with a limit of detection of 0.05 nmol/L (signal-to-noise ratio = 3). The relative standard deviation for six replicate determinations of GSH at the 100 nmol/L concentration level was 3.9%. The method was applied to water samples and average recoveries between 87.5% and 111.5% were obtained for spiked samples.

scholarly journals HPLC Estimation of New Impurity Methyl Ezetimibe in Ezetimibe Drug

2020 ◽  
Vol 32 (6) ◽  
pp. 1309-1313
Author(s):  
Duggirala Parvatha Venkata Vardhani Devi ◽  
Kapavarapu Maruthi Venkata Narayanarao ◽  
Pulipaka Shyamala ◽  
Rallabhandi Murali Krishna ◽  
Komali Siva Prasad
Keyword(s):  
Limit Of Detection ◽  
Signal To Noise Ratio ◽  
Gradient Elution ◽  
Hplc Method ◽  
The Novel ◽  
Signal To Noise ◽  
Drug Substances ◽  
Chromatographic Detection ◽  
Noise Ratio ◽  
Gradient Elution Mode

A new gradient elution mode HPLC method was developed and validated to detect and monitor the novel impurity namely methyl ezitimibe in ezetimibe drug substances. Chromatographic detection and analysis of methyl ezetimibe was performed on XBridge C18 column with mobile phase consisting of 0.02 M phosphate buffer (pH 5) and acetonitrile with 1 mL/min flow rate in gradient elution mode. Methyl ezetimibe was detected and monitored at 248 nm. The calibration curve was linear over range of 0.015 to 0.219% concentration. The limit of detection and quantification were computed as 0.005% (signal to noise ratio 3.60) and 0.015% (signal to noise ratio 15.96), respectively. The precision was 0.97% (%RSD) and accuracy was 93.2 to 98.2% (recovery). The developed method was proved suitable to detect and monitor methyl ezetimibe impurity in ezetimibe drug substances.

Quantitation of Sulfamethazine in Pork Tissue by Thin-Layer Chromatography

1993 ◽  
Vol 76 (2) ◽  
pp. 335-341 ◽  
Author(s):  
Joseph Unruh ◽  
Daniel P Schwartz ◽  
Robert A Barford
Keyword(s):  
Thin Layer ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Signal To Noise Ratio ◽  
Internal Standard ◽  
Signal To Noise ◽  
Thin Layer Chromatographic Plate ◽  
Average Accuracy ◽  
Chromatographic Plate ◽  
Layer Chromatography

Abstract Our earlier method to detect and quantitate sulfamethazine (SMZ) in milk at the 10 ppb level was modified to quantitate SMZ in pork tissue. Sulfabromomethazine (SBZ) is added to the tissue as an internal standard. SMZ and SBZ are extracted from the tissue into water as the supernatant of a centrifuged, aqueous homogenate and are cleaned up and concentrated by a series of solid-phase extractions. The sulfonamide-containing eluate is then separated on a silica gel thin-layer chromatographic plate. SBZ and SMZ are derivatized with fluorescamine, and their fluorescence is quantitated with a scanning densitometer. The limit of detection was estimated at 0.25 ppb (signal-to-noise ratio, 3:1). The average accuracy over the analysis range (0.54-21.8 ppb [μg/kg]) was 95.6% (standard deviation = 29.4%, n = 54).

scholarly journals Digital-resolution detection of microRNA with single-base selectivity by photonic resonator absorption microscopy

2019 ◽  
Vol 116 (39) ◽  
pp. 19362-19367 ◽  
Author(s):  
Taylor D. Canady ◽  
Nantao Li ◽  
Lucas D. Smith ◽  
Yi Lu ◽  
Manish Kohli ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Signal To Noise Ratio ◽  
Resonant Wavelength ◽  
High Signal ◽  
Single Base ◽  
Reflected Light ◽  
Single Base Mismatch ◽  
Base Mismatch ◽  
Digital Resolution ◽  
Target Probe

Circulating exosomal microRNA (miR) represents a new class of blood-based biomarkers for cancer liquid biopsy. The detection of miR at a very low concentration and with single-base discrimination without the need for sophisticated equipment, large volumes, or elaborate sample processing is a challenge. To address this, we present an approach that is highly specific for a target miR sequence and has the ability to provide “digital” resolution of individual target molecules with high signal-to-noise ratio. Gold nanoparticle tags are prepared with thermodynamically optimized nucleic acid toehold probes that, when binding to a target miR sequence, displace a probe-protecting oligonucleotide and reveal a capture sequence that is used to selectively pull down the target-probe–nanoparticle complex to a photonic crystal (PC) biosensor surface. By matching the surface plasmon-resonant wavelength of the nanoparticle tag to the resonant wavelength of the PC nanostructure, the reflected light intensity from the PC is dramatically and locally quenched by the presence of each individual nanoparticle, enabling a form of biosensor microscopy that we call Photonic Resonator Absorption Microscopy (PRAM). Dynamic PRAM imaging of nanoparticle tag capture enables direct 100-aM limit of detection and single-base mismatch selectivity in a 2-h kinetic discrimination assay. The PRAM assay demonstrates that ultrasensitivity (<1 pM) and high selectivity can be achieved on a direct readout diagnostic.

Forward Degenerate Four-Wave Mixing as a Detection Method in Liquid Separation Systems: Improving Detection Limits by Means of a Fabry—Perot Interferometer

1997 ◽  
Vol 51 (7) ◽  
pp. 1008-1011 ◽  
Author(s):  
Gerard Ph. Hoornweg ◽  
Tjipke De Beer ◽  
Nel H. Velthorst ◽  
Cees Gooijer
Keyword(s):  
High Performance ◽  
Limit Of Detection ◽  
Signal To Noise Ratio ◽  
Four Wave Mixing ◽  
Laser Wavelength ◽  
Stray Light ◽  
Concentration Limit ◽  
Wave Mixing ◽  
Fabry Perot ◽  
Degenerate Four Wave Mixing

A simple, small, and low-resolution confocal Fabry–Perot interferometer (CFP) has been developed to improve the signal-to-noise ratio (S/N) in forward degenerate four-wave mixing (F-D4WM) detection, coupled to high-performance liquid chromatography (HPLC). Its appropriateness is based on the difference in coherence between the F-D4WM signal and the background stray light. A detailed description of the specially designed CFP and its performance is presented. The improvement in S/N was calculated from the chromatographic peaks recorded after an HPLC separation of 1- and 2-aminoanthraquinone. The concentration limit of detection (LOD) was improved by a factor of 30; for 1-aminoanthraquinone it was 2 × 10−8 M injected (corresponding to about 3 × 10−9 M in the detector cell), which is quite favorable in view of its low molar extinction coefficient being 2000 M−1 cm−1 at the utilized laser wavelength (514.5 nm).

scholarly journals Physiological Study of Lactobacillus delbrueckii subsp. bulgaricus Strains in a Novel Chemically Defined Medium

2000 ◽  
Vol 66 (12) ◽  
pp. 5306-5311 ◽  
Author(s):  
Christian Chervaux ◽  
S. Dusko Ehrlich ◽  
Emmanuelle Maguin
Keyword(s):  
Carbon Sources ◽  
Streptococcus Thermophilus ◽  
Defined Medium ◽  
Lactobacillus Delbrueckii ◽  
Protein Labeling ◽  
Chemically Defined Medium ◽  
Phosphotransferase System ◽  
Nutritional Conditions ◽  
Labeling Method

ABSTRACT We developed a chemically defined medium called milieu proche du lait (MPL), in which 22 Lactobacillus delbrueckii subsp.bulgaricus (L. bulgaricus) strains exhibited growth rates ranging from 0.55 to 1 h−1. MPL can also be used for cultivation of other lactobacilli and Streptococcus thermophilus. The growth characteristics of L. bulgaricus in MPL containing different carbon sources were determined, including an initial characterization of the phosphotransferase system transporters involved. For the 22 tested strains, growth on lactose was faster than on glucose, mannose, and fructose. Lactose concentrations below 0.4% were limiting for growth. We isolated 2-deoxyglucose-resistant mutants from strains CNRZ397 and ATCC 11842. CNRZ397-derived mutants were all deficient for glucose, fructose, and mannose utilization, indicating that these three sugars are probably transported via a unique mannose-specific-enzyme-II-like transporter. In contrast, mutants of ATCC 11842 exhibited diverse phenotypes, suggesting that multiple transporters may exist in that strain. We also developed a protein labeling method and verified that exopolysaccharide production and phage infection can occur in MPL. The MPL medium should thus be useful in conducting physiological studies ofL. bulgaricus and other lactic acid bacteria under well controlled nutritional conditions.

scholarly journals Eddy-covariance data with low signal-to-noise ratio: time-lag determination, uncertainties and limit of detection

2015 ◽  
Vol 8 (3) ◽  
pp. 2913-2955 ◽  
Author(s):  
B. Langford ◽  
W. Acton ◽  
C. Ammann ◽  
A. Valach ◽  
E. Nemitz
Keyword(s):  
Eddy Covariance ◽  
Random Error ◽  
Limit Of Detection ◽  
Sampling Error ◽  
Signal To Noise Ratio ◽  
Time Lag ◽  
Systematic Bias ◽  
Signal To Noise ◽  
Cross Covariance ◽  
Noise Ratio

Abstract. All eddy-covariance flux measurements are associated with random uncertainties which are a combination of sampling error due to natural variability in turbulence and sensor noise. The former is the principal error for systems where the signal-to-noise ratio of the analyser is high, as is usually the case when measuring fluxes of heat, CO2 or H2O. Where signal is limited, which is often the case for measurements of other trace gases and aerosols, instrument uncertainties dominate. We are here applying a consistent approach based on auto- and cross-covariance functions to quantifying the total random flux error and the random error due to instrument noise separately. As with previous approaches, the random error quantification assumes that the time-lag between wind and concentration measurement is known. However, if combined with commonly used automated methods that identify the individual time-lag by looking for the maximum in the cross-covariance function of the two entities, analyser noise additionally leads to a systematic bias in the fluxes. Combining datasets from several analysers and using simulations we show that the method of time-lag determination becomes increasingly important as the magnitude of the instrument error approaches that of the sampling error. The flux bias can be particularly significant for disjunct data, whereas using a prescribed time-lag eliminates these effects (provided the time-lag does not fluctuate unduly over time). We also demonstrate that when sampling at higher elevations, where low frequency turbulence dominates and covariance peaks are broader, both the probability and magnitude of bias are magnified. We show that the statistical significance of noisy flux data can be increased (limit of detection can be decreased) by appropriate averaging of individual fluxes, but only if systematic biases are avoided by using a prescribed time-lag. Finally, we make recommendations for the analysis and reporting of data with low signal-to-noise and their associated errors.

Novel electrochemiluminescent materials for sensor applications

2014 ◽  
Vol 174 ◽  
pp. 357-367 ◽  
Author(s):  
Lynn Dennany ◽  
Zahera Mohsan ◽  
Alexander L. Kanibolotsky ◽  
Peter J. Skabara
Keyword(s):  
Redox Reactions ◽  
Limit Of Detection ◽  
Signal To Noise Ratio ◽  
High Sensitivity ◽  
Radical Cations ◽  
Detection Sensitivity ◽  
Sensor Development ◽  
Sensor Applications ◽  
Sensor Devices ◽  
The Impact

Electrochemiluminescence (ECL) uses redox reactions to generate light at an electrode surface, and is gaining increasing attention for biosensor development due to its high sensitivity and excellent signal-to-noise ratio. ECL studies of monodisperse oligofluorene–truxenes (T4 series) have been reported previously, showing the production of stable radical cations and radical anions, generating blue ECL. The compound in this study differs from the original structures, in that there are 2,1,3-benzothiadazole (BT) units inserted between the first and second fluorene units of the quarterfluorenyl arms. It was therefore anticipated that the incorporation of these highly luminescent and ECL-active compounds into sensor development would lead to significant decreases in detection limits. In this contribution, we report on the impact of incorporating these novel complexes into sensor devices on the ECL efficiency, as well as the ability of these to improve the detection sensitivity and decrease the limit of detection using the reagent-free detection of model analytes. The real world impact of these compounds is elucidated through the comparison with more standard ECL materials such as ruthenium-based compounds. The potential for multiple applications is to be examined within this contribution.

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