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  • Baldo, BarbaraLund University,Lunds universitet,Translationell neuroendokrinologi,Forskargrupper vid Lunds universitet,Translational Neuroendocrinology,Lund University Research Groups (författare)

Quantification of Total and Mutant Huntingtin Protein Levels in Biospecimens Using a Novel alphaLISA Assay

  • Artikel/kapitelEngelska2018

Förlag, utgivningsår, omfång ...

  • 2018

Nummerbeteckningar

  • LIBRIS-ID:oai:lup.lub.lu.se:5f486647-462e-40a0-a610-5338158cffc8
  • https://lup.lub.lu.se/record/5f486647-462e-40a0-a610-5338158cffc8URI
  • https://doi.org/10.1523/ENEURO.0234-18.2018DOI

Kompletterande språkuppgifter

  • Språk:engelska
  • Sammanfattning på:engelska

Ingår i deldatabas

Klassifikation

  • Ämneskategori:art swepub-publicationtype
  • Ämneskategori:ref swepub-contenttype

Anmärkningar

  • The neurodegenerative Huntington's disease (HD) is caused by a polyglutamine (polyQ) amplification in the huntingtin protein (HTT). Currently there is no effective therapy available for HD; however, several efforts are directed to develop and optimize HTT-lowering methods to improve HD phenotypes. To validate these approaches, there is an immediate need for reliable, sensitive, and easily accessible methods to quantify HTT expression. Using the AlphaLISA platform, we developed two novel sensitive and robust assays for quantification of HTT in biological samples using commercially available antibodies. The first, a polyQ-independent assay, measures the total pool of HTT, while the second, a polyQ-dependent assay, preferentially detects the mutant form of HTT. Using purified HTT protein standards and brain homogenates from an HD mouse model, we determine a lower limit of quantification of 1 and 3 pm and optimal reproducibility with CV values lower than 7% for intra- and 20% for interassay. In addition, we used the assays to quantify HTT in neural stem cells generated from patient-derived induced pluripotent stem cells in vitro and in human brain tissue lysates. Finally, we could detect changes in HTT levels in a mouse model where mutant HTT was conditionally deleted in neural tissue, verifying the potential to monitor the outcome of HTT-lowering strategies. This analytical platform is ideal for high-throughput screens and thus has an added value for the HD community as a tool to optimize novel therapeutic approaches aimed at modulating HTT protein levels.

Ämnesord och genrebeteckningar

Biuppslag (personer, institutioner, konferenser, titlar ...)

  • Sajjad, Muhammad Umar (författare)
  • Cheong, Rachel Y.Lund University,Lunds universitet,Translationell neuroendokrinologi,Forskargrupper vid Lunds universitet,Translational Neuroendocrinology,Lund University Research Groups(Swepub:lu)med-rco (författare)
  • Bigarreau, JulieInstitute For Stem Cell Therapy And Exploration Of Monogenic Diseases (CECS/I-Stem) (författare)
  • Vijayvargia, RaviMassachusetts General Hospital,Harvard University (författare)
  • McLean, CatrionaAlfred Hospital (författare)
  • Perrier, Anselme L.Institute For Stem Cell Therapy And Exploration Of Monogenic Diseases (CECS/I-Stem) (författare)
  • Seong, Ihn SikMassachusetts General Hospital,Harvard University (författare)
  • Halliday, GlendaUniversity of Sydney (författare)
  • Petersén, ÅsaLund University,Lunds universitet,Translationell neuroendokrinologi,Forskargrupper vid Lunds universitet,Translational Neuroendocrinology,Lund University Research Groups(Swepub:lu)mphy-apn (författare)
  • Kirik, DenizLund University,Lunds universitet,Brain Repair and Imaging in Neural Systems (BRAINS),Forskargrupper vid Lunds universitet,Lund University Research Groups(Swepub:lu)mphy-dki (författare)
  • Translationell neuroendokrinologiForskargrupper vid Lunds universitet (creator_code:org_t)

Sammanhörande titlar

  • Ingår i:eNeuro5:42373-2822

Internetlänk

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  • eNeuro (Sök värdpublikationen i LIBRIS)

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