scholarly journals Copper Oxide Nanoparticle-Based Immunosensor for Zearalenone Analysis by Combining Automated Sample Pre-Processing and High-Throughput Terminal Detection

Sensors ◽  
2021 ◽  
Vol 21 (19) ◽  
pp. 6538
Author(s):  
Zhihong Xuan ◽  
Yanxiang Wu ◽  
Hongmei Liu ◽  
Li Li ◽  
Jin Ye ◽  
...  
Keyword(s):  
High Throughput ◽  
Screening Tool ◽  
Detection Method ◽  
Sample Pretreatment ◽  
Fluorescence Signal ◽  
Signal Conversion ◽  
Cuo Nps ◽  
Acidic Conditions ◽  
Oxide Nanoparticle ◽  
Efficient Screening

A rapid and high-throughput fluorescence detection method for zearalenone (ZEN) based on a CuO nanoparticle (NP)-assisted signal amplification immunosensor was developed using an automated sample pretreatment and signal conversion system. CuO NPs with high stability and biocompatibility were used as carriers to immobilize anti-ZEN antibodies. The obtained CuO NP-anti-ZEN can maintain the ability to recognize target toxins and act as both a signal source and carrier to achieve signal conversion using automated equipment. In this process, target toxin detection is indirectly transformed to Cu2+ detection because of the large number of Cu2+ ions released from CuO NPs under acidic conditions. Finally, a simple and high-throughput fluorescence assay based on a fluorescent tripeptide molecule was employed to detect Cu2+, using a multifunctional microporous plate detector. A good linear relationship was observed between the fluorescence signal and the logarithm of ZEN concentration in the range of 16.0–1600.0 μg/kg. Additionally, excellent accuracy with a high recovery yield of 99.2–104.9% was obtained, which was concordant with the results obtained from LC-MS/MS of naturally contaminated samples. The CuO NP-based assay is a powerful and efficient screening tool for ZEN detection and can easily be modified to detect other mycotoxins.

Development of a high-throughput thermoelectric screening tool for combinatorial thin film libraries

2007 ◽  
Vol 254 (3) ◽  
pp. 765-767 ◽  
Author(s):  
M. Otani ◽  
K. Itaka ◽  
W. Wong-Ng ◽  
P.K. Schenck ◽  
H. Koinuma
Keyword(s):  
Thin Film ◽  
High Throughput ◽  
Screening Tool

scholarly journals Determination of Morphine in Human Urine by the Novel Competitive Fluorescence Immunoassay

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Jie Cao ◽  
Xiao-Ying Chen ◽  
Wu-Rong Zhao
Keyword(s):  
Monoclonal Antibodies ◽  
Human Urine ◽  
Screening Tool ◽  
Fluorescein Isothiocyanate ◽  
Cross Reactivity ◽  
The Novel ◽  
Fluorescence Immunoassay ◽  
Coated Plate ◽  
Efficient Screening

A competitive fluorescence immunoassay for the identification and quantification of morphine has been developed on the basis of hapten-coated plate format. Hapten was prepared through covalent conjugating a morphine derivative with albumin bovine. In the immunoassay, the hapten was inoculated on a 96-well plate and then bound with monoclonal antibodies labeled with a signal indicating dye, fluorescein isothiocyanate (FITC). Unbound FITC-antibodies were rinsed off from the plate. The fluorescein intensity decreases in the presence of morphine molecules due to the competitively binding to antibodies against hapten. The intensity is inversely correlated with the concentration of morphine. In quantitative analysis for urine samples, we obtained a linearity range of 0.2 μg/mL∼2.5 μg/mL, along with a detection limit of c.a. 1 ng/mL. The fluorescence immunoassay shows low cross-reactivity (below 10%) to 6-acetylmorphine, 3-acetylmorphine, and heroine. The developed method produced comparable results to the standard GC-MS/MS method. In conclusion, a rapid and efficient screening tool for morphine in clinical human urine has been established.

A Novel High-Throughput Screening Assay for HCN Channel Blocker Using Membrane Potential-Sensitive Dye and FLIPR

2009 ◽  
Vol 14 (9) ◽  
pp. 1119-1128 ◽  
Author(s):  
Dmitry V. Vasilyev ◽  
Qin J. Shan ◽  
Yan T. Lee ◽  
Veronica Soloveva ◽  
Stanley P. Nawoschik ◽  
...  
Keyword(s):  
Membrane Potential ◽  
High Throughput ◽  
High Throughput Screening ◽  
Channel Blocker ◽  
Fluorescent Imaging ◽  
Fluorescence Signal ◽  
Hcn Channel ◽  
Plating Density ◽  
High Throughput Screening Assay ◽  
Cell Plating

Hyperpolarization-activated cation nonselective (HCN) channels represent an interesting group of targets for drug development. In this study, the authors report the development of a novel membrane potential-sensitive dye (MPSD) assay for HCN channel modulators that has been miniaturized into 384-well fluorescent imaging plate reader (FLIPR) high-throughput screening (HTS) format. When optimized (by cell plating density, plate type, cell recovery from cryopreservation), the wellto-well signal variability was low, with a Z' = 0.73 and coefficient of variation = 6.4%, whereas the MPSD fluorescence signal amplitude was -23,700 ± 1500 FLIPR3 relative fluorescence units (a linear relationship was found between HCN1 MPSD fluorescence signal and the cell plating density) and was completely blocked by 30 µM ZD7288. The assay tolerated up to 1% DMSO, inclusion of which did not significantly change the signal kinetics or amplitude. A single-concentration screening of an ion channel-focused library composed of 4855 compounds resulted in 89 HCN1 blocker hits, 51 of which were subsequently analyzed with an 8-point concentration-response analysis on the IonWorks HT electrophysiology platform. The correlation between MPSD and the electrophysiology assay was moderate, as shown by the linear regression analysis (r2 = 0.56) between the respective IC50s obtained using these 2 assays. The reported HTS-compatible HCN channel blocker assay can serve as a tool in drug discovery in the pursuit of HCN channel isoform-selective small molecules that could be used in the development of clinically relevant compounds. (Journal of Biomolecular Screening 2009:1119-1128)

scholarly journals High-throughput Detection Method for Influenza Virus

10.3791/3623 ◽  
2012 ◽  
Author(s):  
Pawan Kumar ◽  
Allison E. Bartoszek ◽  
Thomas M. Moran ◽  
Jack Gorski ◽  
Sanjib Bhattacharyya ◽  
...  
Keyword(s):  
Influenza Virus ◽  
High Throughput ◽  
Detection Method ◽  
High Throughput Detection

High-throughput detection method of quorum-sensing molecules by colorimetry and its applications

2006 ◽  
Vol 356 (2) ◽  
pp. 297-299 ◽  
Author(s):  
Yung-Hun Yang ◽  
Tek-Hyung Lee ◽  
June Hyung Kim ◽  
Eun Jung Kim ◽  
Hwang-Soo Joo ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
High Throughput ◽  
Detection Method ◽  
High Throughput Detection

scholarly journals Chemically diverse polymer microarrays and high throughput surface characterisation: a method for discovery of materials for stem cell culture

2014 ◽  
Vol 2 (11) ◽  
pp. 1604-1611 ◽  
Author(s):  
A. D. Celiz ◽  
J. G. W. Smith ◽  
A. K. Patel ◽  
R. Langer ◽  
D. G. Anderson ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
High Throughput ◽  
Screening Tool ◽  
New Materials ◽  
Surface Characterisation ◽  
Stem Cell Culture

Chemically diverse polymer microarrays as a powerful screening tool for the discovery of new materials for a variety of applications.

scholarly journals Receptor-Ligand Interactions Studied with Homogeneous Fluorescence-Based Assays Suitable for Miniaturized Screening

2001 ◽  
Vol 6 (1) ◽  
pp. 11-18
Author(s):  
Andreas A. Scheel ◽  
Bettina Funsch ◽  
Michael Busch ◽  
Gabriele Gradl ◽  
Johannes Pschorr ◽  
...  
Keyword(s):  
High Throughput ◽  
Single Molecule ◽  
High Throughput Screening ◽  
Free Ligand ◽  
Membrane Vesicles ◽  
Membrane Receptors ◽  
Receptor Expression ◽  
Model Systems ◽  
Fluorescence Signal ◽  
Distribution Analysis

Cell membrane receptors play a central role in controlling cellular functions, making them the target of drugs for a wide variety of diseases. This report describes how a recently developed method, fluorescence intensity distribution analysis (FIDA), can be used to develop homogeneous, nonradioactive high throughput screening assays for membrane receptors. With FIDA, free ligand and ligand accumulated on receptor-bearing membrane vesicles can be distinguished on the basis of their particle brightness. This allows the concentration of both bound and free ligand to be determined reliably from a single measurement, without any separation. We demonstrate that ligand affinity, receptor expression level, and potency of inhibitors can be determined using the epidermal growth factor and β2-adrenergic receptors as model systems. Highly focused confocal optics enable single-molecule sensitivity, and sample volumes can thus be reduced to 1,IL without affecting the quality of the fluorescence signal. Our results demonstrate that FIDA is an ideal method for membrane receptor assays offering substantial benefits for assay development and high throughput pharmaceutical screening.

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