Monoclonal Antibody-Based Fluorescence Polarization Immunoassay for High Throughput Screening of Furaltadone and its Metabolite AMOZ in Animal Feeds and Tissues

2013 ◽  
Vol 16 (6) ◽  
pp. 494-502 ◽  
Author(s):  
Zhen-Lin Xu ◽  
Shi-Wei Zhang ◽  
Yuan-Ming Sun ◽  
Yu-Dong Shen ◽  
Hong-Tao Lei ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Throughput ◽  
Fluorescence Polarization ◽  
High Throughput Screening ◽  
Fluorescence Polarization Immunoassay ◽  
Animal Feeds

Monoclonal Fluorescence Polarization Immunoassay Evaluated for Monitoring Cyclosporine in Whole Blood After Kidney, Heart, and Liver Transplantation

1992 ◽  
Vol 38 (1) ◽  
pp. 123-126 ◽  
Author(s):  
M Winkler ◽  
G Schumann ◽  
D Petersen ◽  
M Oellerich ◽  
K Wonigeit
Keyword(s):  
Monoclonal Antibody ◽  
Liver Transplantation ◽  
Whole Blood ◽  
Fluorescence Polarization ◽  
Fluorescence Polarization Immunoassay ◽  
Transplant Patients ◽  
Specific Radioimmunoassay ◽  
Metabolite Concentrations ◽  
A Prospective Study

Abstract In a prospective study we evaluated a novel fluorescence polarization immunoassay (FPIA) for determining cyclosporine (CsA) in whole blood. FPIA uses a monoclonal antibody and is performed on the TDx (Abbott). The within-series (CV less than 2%) and between-days (CV less than 3.3%) precision of the assay was excellent. The results obtained by the monoclonal FPIA in samples from transplant patients (n = 100) averaged 31.9% and 20.2% higher than those by HPLC and a specific radioimmunoassay (INCStar), respectively. Results by all three methods correlated well. Follow-up studies during the early course after liver transplantation, however, suggested that high metabolite concentrations affect FPIA results. This is explained by previously described cross-reactions of the monoclonal antibody with some CsA metabolites. The FPIA results in samples of such patients should be interpreted cautiously.

The expression of recombinant human LOX-1 and identifying its mimic ligands by fluorescence polarization-based high throughput screening

2006 ◽  
Vol 125 (4) ◽  
pp. 492-502 ◽  
Author(s):  
Tiantai Zhang ◽  
Zhentai Huang ◽  
Ying Dai ◽  
Xiuping Chen ◽  
Ping Zhu ◽  
...  
Keyword(s):  
High Throughput ◽  
Fluorescence Polarization ◽  
High Throughput Screening

Fluorescence polarization immunoassay based on a new monoclonal antibody for the detection of the Diisobutyl phthalate in Yoghurt

Food Control ◽  
2019 ◽  
Vol 105 ◽  
pp. 38-44 ◽  
Author(s):  
Yingshan Chen ◽  
Qiyi He ◽  
Ding Shen ◽  
Zhengyun Jiang ◽  
Sergei A. Eremin ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fluorescence Polarization ◽  
Fluorescence Polarization Immunoassay

scholarly journals A High-Throughput Screening Strategy to Identify Protein-Protein Interaction Inhibitors That Block the Fanconi Anemia DNA Repair Pathway

2016 ◽  
Vol 21 (6) ◽  
pp. 626-633 ◽  
Author(s):  
Andrew F. Voter ◽  
Kelly A. Manthei ◽  
James L. Keck
Keyword(s):  
Dna Repair ◽  
High Throughput ◽  
Fanconi Anemia ◽  
Fluorescence Polarization ◽  
High Throughput Screening ◽  
Titration Calorimetry ◽  
Common Mechanism ◽  
Physical Interaction ◽  
Repair Pathway ◽  
Dna Crosslinking

Induction of the Fanconi anemia (FA) DNA repair pathway is a common mechanism by which tumors evolve resistance to DNA crosslinking chemotherapies. Proper execution of the FA pathway requires interaction between the FA complementation group M protein (FANCM) and the RecQ-mediated genome instability protein (RMI) complex, and mutations that disrupt FANCM/RMI interactions sensitize cells to DNA crosslinking agents. Inhibitors that block FANCM/RMI complex formation could be useful therapeutics for resensitizing tumors that have acquired chemotherapeutic resistance. To identify such inhibitors, we have developed and validated high-throughput fluorescence polarization and proximity assays that are sensitive to inhibitors that disrupt interactions between the RMI complex and its binding site on FANCM (a peptide referred to as MM2). A pilot screen of 74,807 small molecules was performed using the fluorescence polarization assay. Hits from the primary screen were further tested using the proximity assay, and an orthogonal proximity assay was used to assess inhibitor selectivity. Direct physical interaction between the RMI complex and the most selective inhibitor identified through the screening process was measured by surface plasmon resonance and isothermal titration calorimetry. Observation of direct binding by this small molecule validates the screening protocol.

scholarly journals Fluorescence Polarization and Anisotropy in High Throughput Screening: Perspectives and Primer

2000 ◽  
Vol 5 (5) ◽  
pp. 297-306 ◽  
Author(s):  
John C. Owicki
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
Fluorescence Polarization ◽  
High Throughput Screening ◽  
The Past ◽  
Assay Performance ◽  
The Subject ◽  
Homogeneous Assays

Fluorescence polarization and anisotropy are two nearly equivalent techniques that have together, over the past 5 years, achieved wide use in high throughput screening in drug discovery. These are single-label methods that can be used to construct homogeneous assays that are fast, sensitive, and resistant to some significant interferences. Moreover, the assays are relatively inexpensive. This review surveys the peer-reviewed literature on the subject and explores some of the fundamental issues that bear on assay performance.

scholarly journals Fluorescence Polarization Assays for High Throughput Screening of G Protein-Coupled Receptors

2000 ◽  
Vol 5 (3) ◽  
pp. 159-167 ◽  
Author(s):  
Peter Banks ◽  
Mylene Gosselin ◽  
Linda Prystay
Keyword(s):  
High Throughput ◽  
Fluorescence Polarization ◽  
High Throughput Screening ◽  
Rank Order ◽  
Drug Binding ◽  
G Protein Coupled Receptors ◽  
Microtiter Plates ◽  
High Throughput Drug Screening ◽  
G Protein Coupled ◽  
Speed Of Analysis

Fluorescence polarization assays in 384-well microtiter plates have been demonstrated. The performance is suitable for high throughput drug screening applications with respect to speed of analysis, displaceable signal, precision, and sensitivity to various reagents. Rank order of potency was maintained relative to ['251]-ligand filtration assays, and the effects of the highly colored compounds, tartrazine and Chicago Sky Blue, were insignificant on the polarization signal up to a concentration of 1 tiM. These attributes suggest that accurate assessment of drug binding can be obtained.

An improved zinc cocktail-mediated fluorescence polarization-based kinase assay for high-throughput screening of kinase inhibitors

2008 ◽  
Vol 380 (1) ◽  
pp. 143-145 ◽  
Author(s):  
Puneet Chopra ◽  
Kamna Nanda ◽  
Mou Chatterjee ◽  
Malini Bajpai ◽  
Sunanda G. Dastidar ◽  
...  
Keyword(s):  
High Throughput ◽  
Fluorescence Polarization ◽  
High Throughput Screening ◽  
Kinase Inhibitors ◽  
Kinase Assay

scholarly journals Impact of a Red-Shifted Dye Label for High Throughput Fluorescence Polarization Assays of G Protein-Coupled Receptors

2000 ◽  
Vol 5 (5) ◽  
pp. 329-334 ◽  
Author(s):  
Peter Banks ◽  
Mylene Gosselin ◽  
Linda Prystay
Keyword(s):  
G Protein ◽  
High Throughput ◽  
Fluorescence Polarization ◽  
High Throughput Screening ◽  
False Negative ◽  
G Protein Coupled Receptors ◽  
Negative Results ◽  
False Negative Results ◽  
Assay Precision ◽  
G Protein Coupled

High throughput screening fluorescence polarization assays using G protein-coupled receptors (GPCRs) as targets have been compared using fluorescein and BODIPY TMR-labeled peptides. The red-shifted BODIPY TMR dye exhibits improved assay performance relative to fluorescein due to improvement in both ligand affinity to the GPCRs and assay precision brought about by the higher intensity probe. Furthermore, the red-shifted dye demonstrates an insensitivity to the effects of the highly colored compound tartrazine, which can produce false-negative results for assays conducted with fluorescein as a label.

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