Fluorescence Polarization Based Displacement Assay for the Determination of Small Molecules with Aptamers

2008 ◽  
Vol 80 (22) ◽  
pp. 8853-8855 ◽  
Author(s):  
Jorge A. Cruz-Aguado ◽  
Gregory Penner
Keyword(s):  
Small Molecules ◽  
Fluorescence Polarization ◽  
Displacement Assay

2002 W.A.E. McBryde Award Lecture — Affinity recognition, capillary electrophoresis, and laser-induced fluorescence polarization for ultrasensitive bioanalysis

2005 ◽  
Vol 83 (3) ◽  
pp. 185-194 ◽  
Author(s):  
X Chris Le ◽  
Victor Pavski ◽  
Hailin Wang
Keyword(s):  
Dna Damage ◽  
Capillary Electrophoresis ◽  
Small Molecules ◽  
Fluorescence Polarization ◽  
Laser Induced Fluorescence ◽  
Reaction Products ◽  
Wide Range ◽  
Affinity Probes ◽  
Hiv 1

The combination of affinity recognition, capillary electrophoresis (CE), laser-induced fluorescence (LIF), and fluorescence polarization for the ultrasensitive determination of compounds of biological interest is described. Competitive immunoassays using CE–LIF eliminate the need for fluorescently labeling trace analytes of interest and are particularly useful for determination of small molecules, such as cyclosporine, gentamicin, vancomycin, and digoxin. Fluorescence polarization allows for differentiation of the antibody-bound from the unbound small molecules. Noncompetitive affinity CE–LIF assays are shown to be highly effective in the determination of biomarkers for DNA damage and HIV-1 infection. An antibody (or aptamer) is used as a fluorescent probe to bind with a target DNA adduct (or the reverse transcriptase of the HIV-1 virus), with the fluorescent reaction products being separated by CE and detected by LIF. Aptamers are attractive affinity probes for protein analysis because of high affinity, high specificity, and the potential for a wide range of target proteins. Fluorescence polarization provides unique information for studying molecular interactions. Innovative integrations of these technologies will have broad applications ranging from cancer research, to biomedical diagnosis, to pharmaceutical and environmental analyses.Key words: capillary electrophoresis, laser-induced fluorescence, fluorescence polarization, immunoassay, affinity probes, antibodies, aptamers, DNA damage, toxins, therapeutic drugs.

Fluorescence polarization immunoassay for rapid determination of dehydroepiandrosterone in human urine

2021 ◽  
Author(s):  
Huiyi Yang ◽  
Qiyi He ◽  
Sergei A. Eremin ◽  
Junkang Pan ◽  
Yikui Zou ◽  
...  
Keyword(s):  
Human Urine ◽  
Fluorescence Polarization ◽  
Rapid Determination ◽  
Fluorescence Polarization Immunoassay

Development and optimization of a fluorescence polarization immunoassay for orbifloxacin in milk

2014 ◽  
Vol 6 (11) ◽  
pp. 3849-3857 ◽  
Author(s):  
Tiejun Mi ◽  
Xiao Liang ◽  
Long Ding ◽  
Suxia Zhang ◽  
Sergei A. Eremin ◽  
...  
Keyword(s):  
Fluorescence Polarization ◽  
Rapid Determination ◽  
Fluorescence Polarization Immunoassay ◽  
Fluorescent Tracer

Rapid determination of orbifloxacin residue in milk by an optimized fluorescence polarization immunoassay based on a heterogeneous fluorescent tracer.

The determination of neutrophil membrane fluidity in patients with hepatitis B: a fluorescence polarization study

Apmis ◽  
1997 ◽  
Vol 105 (1-6) ◽  
pp. 309-312 ◽  
Author(s):  
XUE-GONG FAN ◽  
ZHENG ZHANG
Keyword(s):  
Hepatitis B ◽  
Membrane Fluidity ◽  
Fluorescence Polarization ◽  
Polarization Study

Working with Small Molecules: Preparing and Storing Stock Solutions and Determination of Kinetic Solubility

2011 ◽  
pp. 265-271 ◽  
Author(s):  
Andrea Wolf ◽  
Satoko Shimamura ◽  
Friedrich B. M. Reinhard
Keyword(s):  
Small Molecules ◽  
Stock Solutions ◽  
Kinetic Solubility

scholarly journals A high‐throughput fluorescence polarization assay for the determination of Kd for soluble epoxide hydrolase inhibitors

2008 ◽  
Vol 22 (S1) ◽  
Author(s):  
Richard H. Ingraham ◽  
Rachel R. Kroe ◽  
Maurice M. Morelock ◽  
John R. Proudfoot ◽  
Anne B. Eldrup ◽  
...  
Keyword(s):  
High Throughput ◽  
Fluorescence Polarization ◽  
Epoxide Hydrolase ◽  
Soluble Epoxide Hydrolase ◽  
Soluble Epoxide Hydrolase Inhibitors ◽  
Fluorescence Polarization Assay

scholarly journals Cryo-EM structure of the NLRP3 decamer bound to the cytokine release inhibitory drug CRID3

2021 ◽  
Author(s):  
Inga V. Hochheiser ◽  
Michael Pilsl ◽  
Gregor Hagelueken ◽  
Jonas Moecking ◽  
Michael Marleaux ◽  
...  
Keyword(s):  
Small Molecules ◽  
Nucleotide Binding Domain ◽  
Native Form ◽  
Spherical Structure ◽  
Cryo Electron Microscopy ◽  
Rational Drug ◽  
Inhibitory Drug ◽  
Drug Optimization ◽  
Apical Axis

NLRP3 is an intracellular sensor protein whose activation by a broad spectrum of exogenous and endogenous stimuli leads to inflammasome formation and pyroptosis. The mechanisms leading to NLRP3 activation and the way how antagonistic small molecules function remain poorly understood. Here we report the cryo-electron microscopy structures of full-length NLRP3 in its native form and complexed with the inhibitor CRID3 (also named MCC950). Inactive, ADP-bound NLRP3 is a decamer composed of homodimers of intertwined LRR domains that assemble back-to-back as pentamers with the NACHT domain located at the apical axis of this spherical structure. Molecular contacts between the concave sites of two opposing LRRs are mediated by an acidic loop extending from an LRR transition segment. Binding of CRID3 significantly stabilizes the NACHT and LRR domains relative to each other, allowing structural resolution of 3.9-4.2 Ang. CRID3 binds into a cleft, connecting four subdomains of the NACHT with the transition LRR. Its central sulfonylurea group interacts with the Walker A motif of the NLRP3 nucleotide-binding domain and is sandwiched between two arginines from opposing sites, explaining the specificity of NLRP3 for this chemical entity. With the determination of the binding site of this lead therapeutic, specific targeting of NLRP3 for the treatment of autoinflammatory and autoimmune diseases and rational drug optimization are within reach.

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