Journal article
Novel monoclonal antibodies against the C-terminal HEAT domain of huntingtin
Journal of Huntington's disease
03/05/2026
DOI: 10.1177/18796397261426634
PMID: 41784089
Abstract
BackgroundReliable detection of huntingtin (HTT) is essential for understanding Huntington's disease (HD) biology and for evaluating therapeutic strategies. However, high-quality monoclonal antibodies (mAbs) against the HTT C-terminal domain remain limited.ObjectiveWe sought to generate and validate novel monoclonal antibodies targeting the HTT C-terminal HEAT-containing domain to better detect HTT independently of potential effects of polyglutamine length that can impact some N-terminally targeted antibodies.MethodsWe immunized mice with a highly purified, well-characterized recombinant protein corresponding to the HTT C-terminal domain. We generated monoclonal antibody-producing hybridoma cell lines and characterized the antibodies in common immuno-applications using parental and HTT-knockout cell lines, and in patient-derived fibroblasts.ResultsThree novel, independent hybridoma lines producing anti-HTT monoclonal antibodies were derived. Using CRISPR-edited HTT knockout cell lines and in patient-derived cells, we identified one clone, anti-HTT [2F8], that was specific and effective across Western blot, immunofluorescence, and ELISA assays. All antibodies bound full-length HTT irrespective of HAP40 interaction or polyQ length in vitro and in cells and showed no cross-reactivity to the N-terminal HEAT domain.ConclusionsThese C-terminal HTT mAbs are thus valuable additional tools for studying endogenous HTT function in both normal and disease contexts.
Details
- Title: Subtitle
- Novel monoclonal antibodies against the C-terminal HEAT domain of huntingtin
- Creators
- Yang-Nim Park - University of Iowa, BiologyRebeka Fanti - University of TorontoSamira Sadeghi - University of TorontoRenu Chandrasekaran - University of TorontoAled M Edwards - Princess Margaret Cancer CentreRachel J Harding - University of TorontoDouglas W Houston - Developmental Studies Hybridoma Bank, Department of Biology, The University of Iowa, 028 BBE, Iowa City, IA 52242-1324, USA
- Resource Type
- Journal article
- Publication Details
- Journal of Huntington's disease
- DOI
- 10.1177/18796397261426634
- PMID
- 41784089
- NLM abbreviation
- J Huntingtons Dis
- ISSN
- 1879-6400
- eISSN
- 1879-6400
- Publisher
- Sage; THOUSAND OAKS
- Grant note
- Natural Sciences and Engineering Research Council of Canada: RGPIN-2024-05769 Schrdinger, LLC: n/a Connaught Fund: n/a NIH/ORIP: R24OD037757 Hereditary Disease Foundation: n/a CIHR: 198025
The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was funded in part by a grant from the NIH/ORIP, R24OD037757 to D.W.H.; R.J.H is supported by funding from CIHR (Funding reference number: 198025), NSERC (RGPIN-2024-05769), CFI, the Hereditary Disease Foundation, Schrodinger LLC, the Connaught Fund, and Conscience - DMOS. R.F. was funded by a MITACS accelerate fellowship. The Structural Genomics Consortium is a registered charity (number: 1097737) that receives funds from Bayer AG, Boehringer Ingelheim, Bristol Myers Squibb, Genentech, Genome Canada through Ontario Genomics Institute [OGI-196], Canada Foundation for Innovation Ontario Research Fund, MITACS, EU/EFPIA/OICR/McGill/KTH/Diamond Innovative Medicines Initiative 2 Joint Undertaking [EUbOPEN grant 875510], Janssen, Merck KGaA (aka EMD in Canada and US), Pfizer, and Takeda.
- Language
- English
- Electronic publication date
- 03/05/2026
- Academic Unit
- Biology
- Record Identifier
- 9985141986302771
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