Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease

Myskiw, Jennifer; Lamoureux, Lise; Peterson, Anne; Knox, David; Jansen, Gerald H.; Coulthart, Michael B.; Booth, Stephanie A.

Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease

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creativework.keywords - en: Creutzfeldt-Jakob disease; Creutzcapillary electrophoresis; Prion
dc.contributor.author: Myskiw, Jennifer; Lamoureux, Lise; Peterson, Anne; Knox, David; Jansen, Gerald H.; Coulthart, Michael B.; Booth, Stephanie A.
dc.date.accessioned: 2023-05-31T16:48:32Z
dc.date.available: 2023-05-31T16:48:32Z
dc.date.issued: 2023-03
dc.description.abstract - en: Creutzfeldt-Jakob disease (CJD) comprises a group of transmissible neurodegenerative diseases with vast phenotypic diversity. Sporadic CJD heterogeneity is predominantly influenced by the genotype at codon 129 of the prion-encoding gene and the molecular weight of PrPSc fragments after protease digestion, resulting in a classification of 6 subtypes of CJD (MM1, MM2, MV1, MV2, VV1, and VV2). The majority of cases with CJD can be distinguished using this classification system. However, a number of reported CJD cases are phenotypically unique from others within their same subtype, such as variably protease-sensitive prionopathies, or exist as a mixture of subtypes within the same patient. Western blotting of brain tissue, along with the genotyping of codon 129 of the prion-encoding gene, is considered the "gold standard" for the biochemical characterization of CJD. Western blotting requires a significant amount of prion protein for detection, is labor-intensive, and is also associated with high interassay variability. In addition to these limitations, a growing body of research suggests that unique subtypes of CJD are often undetected or misdiagnosed using standard diagnostic western blotting protocols. Consequently, we successfully optimized and developed a capillary-based western assay using the JESS Simple Western (ProteinSimple) to detect and characterize prion proteins from patients with CJD. We found that this novel assay consistently differentiated CJD type 1 and type 2 cases with a limit of detection 10 to 100× higher than traditional western blotting. Cases with CJD in which type 1 and type 2 coexist within the same brain region can be detected using type 1-specific and type 2-specific antibodies, and we found that there was remarkable specificity for the detection of cases with variably protease-sensitive prionopathy. The assay presented displays outstanding sensitivity, allowing for the preservation of valuable samples and enhancing current detection methods.
dc.identifier.citation: Myskiw J, Lamoureux L, Peterson A, Knox D, Jansen GH, Coulthart MB, Booth SA. Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease. Lab Invest. 2023 Mar;103(3):100029. doi: 10.1016/j.labinv.2022.100029
dc.identifier.doi: https://doi.org/10.1016/j.labinv.2022.100029
dc.identifier.issn: 1530-0307
dc.identifier.uri: https://open-science.canada.ca/handle/123456789/461
dc.language.iso: en
dc.publisher: Nature
dc.rights - en: Open Government Licence - Canada
dc.rights - fr: Licence du gouvernement ouvert - Canada
dc.rights.openaccesslevel - en: Green
dc.rights.openaccesslevel - fr: Vert
dc.rights.uri - en: https://open.canada.ca/en/open-government-licence-canada
dc.rights.uri - fr: https://ouvert.canada.ca/fr/licence-du-gouvernement-ouvert-canada
dc.subject - en: Health
dc.subject - fr: Santé
dc.subject.en - en: Health
dc.subject.fr - fr: Santé
dc.title - en: Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease
dc.type - en: Article
dc.type - fr: Article
local.article.journalissue: 3
local.article.journaltitle: Laboratory Investigation
local.article.journalvolume: 103
local.peerreview - en: Yes
local.peerreview - fr: Oui