{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T19:29:22Z","timestamp":1790278162561,"version":"4.1.0"},"reference-count":19,"publisher":"SAGE Publications","issue":"3","license":[{"start":{"date-parts":[[2022,8,30]],"date-time":"2022-08-30T00:00:00Z","timestamp":1661817600000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JHD"],"published-print":{"date-parts":[[2022,8,30]]},"abstract":"<jats:p>Background: The use of biomarkers has become a major component of clinical trial design. In Huntington\u2019s disease (HD), quantifying the amount of huntingtin protein (HTT) in patient cerebrospinal fluid (CSF) has served as a pharmacodynamic readout for HTT-lowering therapeutic approaches and is a potential disease progression biomarker. To date, an ultrasensitive immunoassay to quantify mutant HTT protein (mHTT) has been used, but additional assays are needed to measure other forms of HTT protein. Objective: We aimed to develop an ultrasensitive immunoassay to quantify HTT protein in a polyglutamine length-independent manner (mHTT and non-expanded wild type HTT combined) in control and HD participant CSF samples. Methods: An ultrasensitive, bead-based, single molecule counting (SMC) immunoassay platform was used for the detection of HTT protein in human CSF samples. Results: A novel ultrasensitive SMC immunoassay was developed to quantify HTT protein in a polyglutamine length-independent manner and shown to measure HTT in both control and HD participant CSF samples. We validate the selectivity and specificity of the readout using biochemical and molecular biology tools, and we undertook a preliminary analytical qualification of this assay to enable its clinical use. We also used this novel assay, along with the previously described mHTT assay, to analyze CSF from control and HD participants. The results of this preliminary set suggests that correlation is present between mHTT and the polyglutamine length-independent HTT levels in human CSF. Conclusion: We have developed a novel ultrasensitive immunoassay that is able to quantify HTT protein in a polyglutamine length-independent manner in control and HD participant CSF.<\/jats:p>","DOI":"10.3233\/jhd-220527","type":"journal-article","created":{"date-parts":[[2022,8,5]],"date-time":"2022-08-05T11:22:42Z","timestamp":1659698562000},"page":"291-305","source":"Crossref","is-referenced-by-count":11,"title":["Quantifying Huntingtin Protein in Human Cerebrospinal Fluid Using a Novel Polyglutamine Length-Independent Assay"],"prefix":"10.1177","volume":"11","author":[{"given":"Valentina","family":"Fodale","sequence":"first","affiliation":[{"name":"Department of Translational and Discovery Research, IRBM S.p.A., Pomezia, Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberta","family":"Pintauro","sequence":"additional","affiliation":[{"name":"Department of Translational and Discovery Research, IRBM S.p.A., Pomezia, Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manuel","family":"Daldin","sequence":"additional","affiliation":[{"name":"Department of Translational and Discovery Research, IRBM S.p.A., Pomezia, Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Carolina","family":"Spiezia","sequence":"additional","affiliation":[{"name":"Department of Translational and Discovery Research, IRBM S.p.A., Pomezia, Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas","family":"Macdonald","sequence":"additional","affiliation":[{"name":"CHDI Management\/CHDI Foundation, Los Angeles, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alberto","family":"Bresciani","sequence":"additional","affiliation":[{"name":"Department of Translational and Discovery Research, IRBM S.p.A., Pomezia, Roma, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","reference":[{"key":"10.3233\/JHD-220527_ref1","first-page":"971","article-title":"The Huntington\u2019s Disease Collaborative Research Group","volume":"72","author":"The Huntington\u2019s Disease Collaborative Research GrouA novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington\u2019s disease chromosomes","year":"1993","journal-title":"Cell"},{"key":"10.3233\/JHD-220527_ref2","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1055\/s-2008-1040853","article-title":"Recent insights into the molecular pathogenesis of Huntington disease","volume":"19","author":"Leavitt","year":"1999","journal-title":"Semin Neurol"},{"key":"10.3233\/JHD-220527_ref3","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1002\/mds.26020","article-title":"Huntingtin-lowering strategies in Huntington\u2019s disease: Antisense oligonucleotides, small RNAs, and gene editing","volume":"29","author":"Aronin","year":"2014","journal-title":"Mov Disord"},{"key":"10.3233\/JHD-220527_ref4","first-page":"6","article-title":"Ultrasensitive measurement of huntingtin protein in cerebrospinal fluid demonstrates increase with Huntington disease stage and decrease following brain huntingtin suppression","volume":"5","author":"Southwell","year":"1216","journal-title":"Sci Rep"},{"key":"10.3233\/JHD-220527_ref5","doi-asserted-by":"crossref","first-page":"1979","DOI":"10.1172\/JCI80743","article-title":"Quantification of mutant huntingtin protein in cerebrospinal fluid from Huntington\u2019s disease patients","volume":"125","author":"Wild","year":"2015","journal-title":"J Clin Invest"},{"key":"10.3233\/JHD-220527_ref6","doi-asserted-by":"crossref","first-page":"349","DOI":"10.3233\/JHD-170269","article-title":"Validation of ultrasensitive mutant huntingtin detection in human cerebrospinal fluid by single molecule counting immunoassay","volume":"6","author":"Fodale","year":"2017","journal-title":"J Huntingtons Dis"},{"key":"10.3233\/JHD-220527_ref8","doi-asserted-by":"crossref","first-page":"e112262","DOI":"10.1371\/journal.pone.0112262","article-title":"Polyglutamine- and temperature-dependent conformational rigidity in mutant huntingtin revealed by immunoassays and circular dichroism spectroscopy","volume":"9","author":"Fodale","year":"2014","journal-title":"PLoS One"},{"key":"10.3233\/JHD-220527_ref9","doi-asserted-by":"crossref","first-page":"22137","DOI":"10.1038\/s41598-020-78790-5","article-title":"Analysis of mutant and total huntingtin expression in Huntington\u2019s disease murine models","volume":"10","author":"Fodale","year":"2020","journal-title":"Sci Rep"},{"key":"10.3233\/JHD-220527_ref10","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1523\/JNEUROSCI.1865-20.2020","article-title":"Mutant huntingtin is cleared from the brain via active mechanisms in Huntington 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