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Fluorescent Filter-Trap Assay for Amyloid Fibril Formation Kinetics in Complex Solutions.

Nasir, Irem LU ; Linse, Sara LU and Cabaleiro-Lago, Celia LU (2015) In ACS Chemical Neuroscience 6(8). p.1436-1444
Abstract
Amyloid fibrils are the most distinct components of the plaques associated with various neurodegenerative diseases. Kinetic studies of amyloid fibril formation shed light on the microscopic mechanisms that underlie this process as well as the contributions of internal and external factors to the interplay between different mechanistic steps. Thioflavin T is a widely used noncovalent fluorescent probe for monitoring amyloid fibril formation; however, it may suffer from limitations due to the unspecific interactions between the dye and the additives. Here, we present the results of a filter-trap assay combined with the detection of fluorescently labeled amyloid β (Aβ) peptide. The filter-trap assay separates formed aggregates based on size,... (More)
Amyloid fibrils are the most distinct components of the plaques associated with various neurodegenerative diseases. Kinetic studies of amyloid fibril formation shed light on the microscopic mechanisms that underlie this process as well as the contributions of internal and external factors to the interplay between different mechanistic steps. Thioflavin T is a widely used noncovalent fluorescent probe for monitoring amyloid fibril formation; however, it may suffer from limitations due to the unspecific interactions between the dye and the additives. Here, we present the results of a filter-trap assay combined with the detection of fluorescently labeled amyloid β (Aβ) peptide. The filter-trap assay separates formed aggregates based on size, and the fluorescent label attached to Aβ allows for their detection. The times of half completion of the process (t1/2) obtained by the filter-trap assay are comparable to values from the ThT assay. High concentrations of human serum albumin (HSA) and carboxyl-modified polystyrene nanoparticles lead to an elevated ThT signal, masking a possible fibril formation event. The filter-trap assay allows fibril formation to be studied in the presence of those substances and shows that Aβ fibril formation is kinetically inhibited by HSA and that the amount of fibrils formed are reduced. In contrast, nanoparticles exhibit a dual-behavior governed by their concentration. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
ACS Chemical Neuroscience
volume
6
issue
8
pages
1436 - 1444
publisher
The American Chemical Society
external identifiers
  • pmid:25946560
  • wos:000359967300020
  • scopus:84939782105
ISSN
1948-7193
DOI
10.1021/acschemneuro.5b00104
language
English
LU publication?
yes
id
85de848a-9cc9-426a-833a-082ff30a47ff (old id 5456849)
date added to LUP
2015-06-23 17:33:36
date last changed
2017-10-01 04:21:52
@article{85de848a-9cc9-426a-833a-082ff30a47ff,
  abstract     = {Amyloid fibrils are the most distinct components of the plaques associated with various neurodegenerative diseases. Kinetic studies of amyloid fibril formation shed light on the microscopic mechanisms that underlie this process as well as the contributions of internal and external factors to the interplay between different mechanistic steps. Thioflavin T is a widely used noncovalent fluorescent probe for monitoring amyloid fibril formation; however, it may suffer from limitations due to the unspecific interactions between the dye and the additives. Here, we present the results of a filter-trap assay combined with the detection of fluorescently labeled amyloid β (Aβ) peptide. The filter-trap assay separates formed aggregates based on size, and the fluorescent label attached to Aβ allows for their detection. The times of half completion of the process (t1/2) obtained by the filter-trap assay are comparable to values from the ThT assay. High concentrations of human serum albumin (HSA) and carboxyl-modified polystyrene nanoparticles lead to an elevated ThT signal, masking a possible fibril formation event. The filter-trap assay allows fibril formation to be studied in the presence of those substances and shows that Aβ fibril formation is kinetically inhibited by HSA and that the amount of fibrils formed are reduced. In contrast, nanoparticles exhibit a dual-behavior governed by their concentration.},
  author       = {Nasir, Irem and Linse, Sara and Cabaleiro-Lago, Celia},
  issn         = {1948-7193},
  language     = {eng},
  number       = {8},
  pages        = {1436--1444},
  publisher    = {The American Chemical Society},
  series       = {ACS Chemical Neuroscience},
  title        = {Fluorescent Filter-Trap Assay for Amyloid Fibril Formation Kinetics in Complex Solutions.},
  url          = {http://dx.doi.org/10.1021/acschemneuro.5b00104},
  volume       = {6},
  year         = {2015},
}