Fluorescent Filter-Trap Assay for Amyloid Fibril Formation Kinetics in Complex Solutions.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/5456849
- author
- Nasir, Irem LU ; Linse, Sara LU and Cabaleiro-Lago, Celia LU
- organization
- publishing date
- 2015
- type
- Contribution to journal
- publication status
- published
- subject
- in
- ACS Chemical Neuroscience
- volume
- 6
- issue
- 8
- pages
- 9 pages
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- pmid:25946560
- wos:000359967300020
- scopus:84939782105
- pmid:25946560
- 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
- 2016-04-01 14:23:20
- date last changed
- 2023-09-03 13:45:09
@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 (ACS)}},
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}},
doi = {{10.1021/acschemneuro.5b00104}},
volume = {{6}},
year = {{2015}},
}