You are here

FAU Collections » FAU Research Repository » FAU College Collections

A Kinetic Aggregation Assay Enabling Selective and Sensitive Aβ Amyloid Quantification in Cells and Tissues

Download pdf | Full Screen View

Date Issued:
2011
Summary:
The process of amyloid-β (Aβ) fibril formation is genetically and pathologically linked to Alzheimer's disease (AD). Thus, a selective and sensitive method for the quantification of Aβ amyloid fibrils in complex biological samples enables a variety of hypotheses to be tested. Herein we report the basis for a quantitative in vitro kinetic aggregation assay that detects seeding competent Aβ aggregates in mammalian cell culture media, in Caenorhabditis elegans lysate and in mouse brain homogenate. Sonicated, proteinase K treated Aβ-fibril-containing tissue homogenates or cell culture media were added to an initially monomeric Aβ1–40 reporter peptide to seed an in vitro nucleated aggregation reaction. The reduction in the half time (t50) of the amyloid growth phase is proportional to the quantity of seeding-competent Aβ aggregates present in the biological sample. An ion exchange resin amyloid isolation strategy from complex biological samples is demonstrated as an alternative to improve the sensitivity and linearity of the kinetic aggregation assay.
Title: A Kinetic Aggregation Assay Enabling Selective and Sensitive Aβ Amyloid Quantification in Cells and Tissues.
34 views
16 downloads
Name(s): Deguo Du
Amber N. Murray
Ehud Cohen
Hyun-Eui Kim
Ryan Simkovsky
Andrew Dillin
Jeffery W. Kelly
Type of Resource: text
Date Issued: 2011
Physical Form: pdf
Extent: 21 p.
Language(s): English
Summary: The process of amyloid-β (Aβ) fibril formation is genetically and pathologically linked to Alzheimer's disease (AD). Thus, a selective and sensitive method for the quantification of Aβ amyloid fibrils in complex biological samples enables a variety of hypotheses to be tested. Herein we report the basis for a quantitative in vitro kinetic aggregation assay that detects seeding competent Aβ aggregates in mammalian cell culture media, in Caenorhabditis elegans lysate and in mouse brain homogenate. Sonicated, proteinase K treated Aβ-fibril-containing tissue homogenates or cell culture media were added to an initially monomeric Aβ1–40 reporter peptide to seed an in vitro nucleated aggregation reaction. The reduction in the half time (t50) of the amyloid growth phase is proportional to the quantity of seeding-competent Aβ aggregates present in the biological sample. An ion exchange resin amyloid isolation strategy from complex biological samples is demonstrated as an alternative to improve the sensitivity and linearity of the kinetic aggregation assay.
Identifier: FAUIR000498 (IID)
Persistent Link to This Record: http://purl.flvc.org/fau/fd/FAUIR000498
Host Institution: FAU

In Collections