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A11-positive β-amyloid Oligomer Preparation and Assessment Using Dot Blotting Analysis
Published on: May 22, 2018
Structure-selective anisotropy assay for amyloid Beta oligomers
Evgenia G Matveeva1, Alan Rudolph, Jonathan R Moll
1University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, Baltimore, Maryland 21201, United States; Adlyfe, Inc., Rockville, Maryland 20850, United States. evgenia_matveeva@hotmail.com
ACS Chemical Neuroscience
|November 28, 2012
Summary
A new fluorescence assay specifically detects toxic amyloid beta (Abeta) oligomers in Alzheimer's disease (AD) research. This method aids in measuring Abeta oligomers in biological samples like cerebrospinal fluid.
Area of Science:
- Neuroscience
- Biochemistry
- Analytical Chemistry
Background:
- Oligomeric amyloid beta (Abeta) species are implicated as key neurotoxic agents in Alzheimer's disease (AD).
- Detecting and quantifying these specific oligomeric forms in biological samples presents a significant challenge.
- Existing methods often lack specificity for oligomers over monomers or fibrils.
Purpose of the Study:
- To develop a novel, oligomer-specific assay for detecting amyloid beta (Abeta) oligomers.
- To provide a tool for accurate measurement of Abeta oligomers in biological fluids such as cerebrospinal fluid (CSF).
- To facilitate further research into the role of Abeta oligomers in AD pathogenesis.
Main Methods:
- Development of a conformationally sensitive, bis-pyrene-labeled peptide (pronucleon peptide, PP) targeting Abeta residues 16-35.
- Utilizing a fluorescence assay that monitors changes in pyrene emission (excimer to monomer) upon binding to Abeta oligomers.
- Employing fluorescence anisotropy (polarization) measurements to specifically detect Abeta oligomer binding in solution.
Main Results:
- The PP assay demonstrates specificity for Abeta oligomers, distinguishing them from monomers and fibrils.
- Binding of PP to Abeta oligomers induces a measurable shift in fluorescence emission spectra.
- Changes in fluorescence anisotropy provide a sensitive readout for Abeta oligomer detection.
Conclusions:
- A simple, rapid fluorescence anisotropy assay for Abeta oligomers has been successfully developed.
- This assay enables specific detection and measurement of Abeta oligomers in biological samples.
- The assay is a valuable tool for advancing Alzheimer's disease research and diagnostics.
