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

被引：3

作者：

Fischer, David F. ^{[1
]}

Dijkstra, Sipke ^{[2
]}

Lo, Kimberly ^{[2
]}

Suijker, Johnny ^{[2
,6
]}

Correia, Ana C. P. ^{[2
]}

Naud, Patricia ^{[3
]}

Poirier, Martin ^{[3
,7
]}

Tessari, Michela A. ^{[4
]}

Boogaard, Ivette ^{[2
]}

Flynn, Geraldine ^{[2
]}

Visser, Mijke ^{[2
]}

Lamers, Marieke B. A. C. ^{[1
]}

McAllister, George ^{[1
,5
]}

Munoz-Sanjuan, Ignacio ^{[5
]}

Macdonald, Douglas ^{[5
]}

机构：

[1] Charles River, Chesterford Res Pk, Saffron Walden, Essex, England

[2] Charles River, Leiden, Netherlands

[3] Charles River, Shrewsbury, MA USA

[4] Galapagos, Leiden, Netherlands

[5] CHDI Fdn, CHDI Management, Los Angeles, CA 90045 USA

[6] Mimetas, Leiden, Netherlands

[7] Altasciences, Everett, WA USA

来源：

PLOS ONE | 2022年 / 17卷 / 04期

关键词：

IN MOUSE MODEL; MUTANT HUNTINGTIN; CEREBROSPINAL-FLUID; CAG REPEAT; DISEASE; PROTEIN; CONFORMATION; SUPPRESSION; EXPANSION; REVEALS;

D O I：

10.1371/journal.pone.0266812

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：20