Optimization of an HTRF Assay for the Detection of Soluble Mutant Huntingtin in Human Buffy Coats: A Potential Biomarker in Blood for Huntington Disease

A means for measuring levels of soluble huntingtin proteins in clinical samples is essential for assessing the biological effects of potential mutant huntingtin (mtHtt) modifying treatments being developed for Huntington's disease (HD). We have optimized a previously described cell-based Homoge...

Full description

Saved in:
Bibliographic Details
Published in:PLoS currents 2010-12, Vol.2, p.RRN1205-RRN1205
Main Authors: Moscovitch-Lopatin, Miriam, Weiss, Andreas, Rosas, Herminia Diana, Ritch, James, Doros, Gheorghe, Kegel, Kimberly B, Difiglia, Marian, Kuhn, Rainer, Bilbe, Graeme, Paganetti, Paolo, Hersch, Steven
Format: Article
Language:English
Subjects:
Huntington Disease
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A means for measuring levels of soluble huntingtin proteins in clinical samples is essential for assessing the biological effects of potential mutant huntingtin (mtHtt) modifying treatments being developed for Huntington's disease (HD). We have optimized a previously described cell-based Homogeneous Time Resolved Fluorescence method that can measure soluble mtHtt and its ratio to the total Htt (tHtt) in blood buffy coats [1]. The results of the optimization and assay qualification indicate the assay to be specific for mtHtt in HD compared to Control subjects, highly sensitive, and technically and biologically reproducible. We therefore generated a Good Laboratory Practice Standard Operating Procedure which we validated, using 30 HD and 8 control buffy coat samples in which significant differences in mtHtt levels were found. We intend to deploy the assay to evaluate sample sets from observational and therapeutic studies enrolling HD subjects to further validate soluble mtHtt measurement by HTRF as a biomarker for HD and to explore its potential uses.
ISSN:2157-3999
2157-3999
DOI:10.1371/currents.rrn1205