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

PLoS ONE, vol. 17, pp. e0266812

Abstract

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.

Authors 12

1. David F. Fischer corresponding
   Affiliation as printed

   Charles River, Chesterford Research Park, Saffron Walden, United Kingdom

2. Sipke Dijkstra corresponding Aachen Charles River

   Charles River Laboratories (Netherlands)

   Affiliation as printed

   Charles River, Leiden, The Netherlands

3. Kimberly Lo corresponding Aachen Charles River

   Charles River Laboratories (Netherlands)

   Affiliation as printed

   Charles River, Leiden, The Netherlands

4. Johnny Suijker corresponding Aachen Charles River

   Charles River Laboratories (Netherlands)

   Affiliation as printed

   Charles River, Leiden, The Netherlands

5. Ana C. P. Correia corresponding Aachen Charles River

   Charles River Laboratories (Netherlands)

   Affiliation as printed

   Charles River, Leiden, The Netherlands

6. Patricia Naud corresponding
   Affiliation as printed

   Charles River, Shrewsbury, MA, United States of America

7. Ivette Boogaard corresponding Aachen Charles River

   Charles River Laboratories (Netherlands)

   Affiliation as printed

   Charles River, Leiden, The Netherlands

8. Mijke Visser corresponding Aachen Charles River

   Charles River Laboratories (Netherlands)

   Affiliation as printed

   Charles River, Leiden, The Netherlands

9. Marieke B. A. C. Lamers corresponding
   Affiliation as printed

   Charles River, Chesterford Research Park, Saffron Walden, United Kingdom

10. George McAllister corresponding

    CHDI Foundation

    Affiliation as printed

    CHDI Management/CHDI Foundation, Los Angeles, CA, United States of America

    Charles River, Chesterford Research Park, Saffron Walden, United Kingdom

11. Ignacio Muñoz-Sanjuán corresponding

    CHDI Foundation

    Affiliation as printed

    CHDI Management/CHDI Foundation, Los Angeles, CA, United States of America

12. Douglas Macdonald corresponding

    CHDI Foundation

    Affiliation as printed

    CHDI Management/CHDI Foundation, Los Angeles, CA, United States of America