title: Optimization of an HTRF Assay for the Detection of Soluble Mutant Huntingtin in Human Buffy Coats: A Potential Biomarker in Blood for Huntington Disease.
abstract: 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.
authors.affiliation: Massachusetts General Hospital/Harvard Medical School, Boston, Massachusetts; Novartis Institutes for Biomedical Research, Neuroscience; Boston University; Massachusetts General Hospital/Harvard Medical School; Massachusetts General Hospital; Neuroscience Research, Head Neurodegeneration at Novartis, Basel, Switzerland; Novartis Institutes for Biomedical Research and AC Immune SA.
authors.forename: Miriam
authors.initials: M
authors.lastname: Moscovitch-Lopatin
authors.forename: Andreas
authors.initials: A
authors.lastname: Weiss
authors.forename: Herminia Diana
authors.initials: HD
authors.lastname: Rosas
authors.forename: James
authors.initials: J
authors.lastname: Ritch
authors.forename: Gheorghe
authors.initials: G
authors.lastname: Doros
authors.forename: Kimberly B
authors.initials: KB
authors.lastname: Kegel
authors.forename: Marian
authors.initials: M
authors.lastname: Difiglia
authors.forename: Rainer
authors.initials: R
authors.lastname: Kuhn
authors.forename: Graeme
authors.initials: G
authors.lastname: Bilbe
authors.forename: Paolo
authors.initials: P
authors.lastname: Paganetti
authors.forename: Steven
authors.initials: S
authors.lastname: Hersch