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

Miriam MoscovitchLopatin | Zendy; Andreas Weiss | Zendy; H. Diana Rosas | Zendy; James J. Ritch | Zendy; Gheorghe Doros | Zendy; Kimberly B. Kegel | Zendy; Marian DiFiglia | Zendy; Rainer Kühn | Zendy; Graeme Bilbe | Zendy; Paolo Paganetti | Zendy; Steven M. Hersch | Zendy

Open Access

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

Author(s) -

Miriam MoscovitchLopatin,

Andreas Weiss,

H. Diana Rosas,

James J. Ritch,

Gheorghe Doros,

Kimberly B. Kegel,

Marian DiFiglia,

Rainer Kühn,

Graeme Bilbe,

Paolo Paganetti,

Steven M. Hersch

Publication year - 2010

Publication title -

plos currents

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.282

H-Index - 49

ISSN - 2157-3999

DOI - 10.1371/currents.rrn1205

Subject(s) - huntingtin , buffy coat , huntington's disease , mutant , biomarker , homogeneous , biology , computational biology , microbiology and biotechnology , medicine , biochemistry , immunology , disease , pathology , gene , thermodynamics , physics

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.

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