Validation of Ultrasensitive Mutant Huntingtin Detection in Human Cerebrospinal Fluid by Single Molecule Counting Immunoassay.

Autor: Fodale V; IRBM Science Park, Pomezia, Rome, Italy.; IRBM Promidis, Pomezia, Rome, Italy., Boggio R; IRBM Promidis, Pomezia, Rome, Italy., Daldin M; IRBM Science Park, Pomezia, Rome, Italy., Cariulo C; IRBM Science Park, Pomezia, Rome, Italy., Spiezia MC; IRBM Science Park, Pomezia, Rome, Italy., Byrne LM; University College London (UCL) Institute of Neurology, National Hospital for Neurology and Neurosurgery, London, UK., Leavitt BR; Department of Medical Genetics, Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, BC, Canada., Wild EJ; University College London (UCL) Institute of Neurology, National Hospital for Neurology and Neurosurgery, London, UK., Macdonald D; CHDIManagement/CHDI Foundation, Los Angeles, CA, USA., Weiss A; IRBM Promidis, Pomezia, Rome, Italy., Bresciani A; IRBM Science Park, Pomezia, Rome, Italy.
Jazyk: angličtina
Zdroj: Journal of Huntington's disease [J Huntingtons Dis] 2017; Vol. 6 (4), pp. 349-361.
DOI: 10.3233/JHD-170269
Abstrakt: Background: The measurement of disease-relevant biomarkers has become a major component of clinical trial design, but in the absence of rigorous clinical and analytical validation of detection methodology, interpretation of results may be misleading. In Huntington's disease (HD), measurement of the concentration of mutant huntingtin protein (mHTT) in cerebrospinal fluid (CSF) of patients may serve as both a disease progression biomarker and a pharmacodynamic readout for HTT-lowering therapeutic approaches. We recently published the quantification of mHTT levels in HD patient CSF by a novel ultrasensitive immunoassay-based technology and here analytically validate it for use.
Objective: This work aims to analytically and clinically validate our ultrasensitive assay for mHTT measurement in human HD CSF, for application as a pharmacodynamic biomarker of CNS mHTT lowering in clinical trials.
Methods: The single molecule counting (SMC) assay is an ultrasensitive bead-based immunoassay where upon specific recognition, dye-labeled antibodies are excited by a confocal laser and emit fluorescent light as a readout. The detection of mHTT by this technology was clinically validated following established Food and Drug Administration and European Medicine Agency guidelines.
Results: The SMC assay was demonstrated to be accurate, precise, specific, and reproducible. While no matrix influence was detected, a list of interfering substances was compiled as a guideline for proper collection and storage of patient CSF samples. In addition, a set of recommendations on result interpretation is provided.
Conclusions: This SMC assay is a robust and ultrasensitive method for the relative quantification of mHTT in human CSF.
Databáze: MEDLINE
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