Development of an Automated Capillary Immunoassay to Detect Prion Glycotypes in Creutzfeldt-Jakob Disease
- DOI
- Language of the publication
- English
- Date
- 2023-03
- Type
- Article
- Author(s)
- Myskiw, Jennifer
- Lamoureux, Lise
- Peterson, Anne
- Knox, David
- Jansen, Gerald H.
- Coulthart, Michael B.
- Booth, Stephanie A.
- Publisher
- Nature
Abstract
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.
Subject
- Health
Keywords
- Creutzfeldt-Jakob disease,
- Creutzcapillary electrophoresis,
- Prion
Rights
Peer review
Yes
Open access level
Green
Identifiers
- ISSN
- 1530-0307
Article
- Journal title
- Laboratory Investigation
- Journal volume
- 103
- Journal issue
- 3
Citation(s)
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