Development of mAb-based polyglutamine-dependent and polyglutamine length-independent huntingtin quantification assays with cross-site validation.

Autor: Fischer DF; Charles River, Chesterford Research Park, Saffron Walden, United Kingdom., Dijkstra S; Charles River, Leiden, The Netherlands., Lo K; Charles River, Leiden, The Netherlands., Suijker J; Charles River, Leiden, The Netherlands., Correia ACP; Charles River, Leiden, The Netherlands., Naud P; Charles River, Shrewsbury, MA, United States of America., Poirier M; Charles River, Shrewsbury, MA, United States of America., Tessari MA; Galapagos, Leiden, The Netherlands., Boogaard I; Charles River, Leiden, The Netherlands., Flynn G; Charles River, Leiden, The Netherlands., Visser M; Charles River, Leiden, The Netherlands., Lamers MBAC; Charles River, Chesterford Research Park, Saffron Walden, United Kingdom., McAllister G; Charles River, Chesterford Research Park, Saffron Walden, United Kingdom.; CHDI Management/CHDI Foundation, Los Angeles, CA, United States of America., Munoz-Sanjuan I; CHDI Management/CHDI Foundation, Los Angeles, CA, United States of America., Macdonald D; CHDI Management/CHDI Foundation, Los Angeles, CA, United States of America.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2022 Apr 08; Vol. 17 (4), pp. e0266812. Date of Electronic Publication: 2022 Apr 08 (Print Publication: 2022).
DOI: 10.1371/journal.pone.0266812
Abstrakt: Huntington's disease (HD) is caused by an expansion of the CAG trinucleotide repeat domain in the huntingtin gene that results in expression of a mutant huntingtin protein (mHTT) containing an expanded polyglutamine tract in the amino terminus. A number of therapeutic approaches that aim to reduce mHTT expression either locally in the CNS or systemically are in clinical development. We have previously described sensitive and selective assays that measure human HTT proteins either in a polyglutamine-independent (detecting both mutant expanded and non-expanded proteins) or in a polyglutamine length-dependent manner (detecting the disease-causing polyglutamine repeats) on the electrochemiluminescence Meso Scale Discovery detection platform. These original assays relied upon polyclonal antibodies. To ensure an accessible and sustainable resource for the HD field, we developed similar assays employing monoclonal antibodies. We demonstrate that these assays have equivalent sensitivity compared to our previous assays through the evaluation of cellular and animal model systems, as well as HD patient biosamples. We also demonstrate cross-site validation of these assays, allowing direct comparison of studies performed in geographically distinct laboratories.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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