1. An Aggregation-Specific Enzyme-Linked Immunosorbent Assay: Detection of Conformational Differences between Recombinant PrP Protein Dimers and PrP Sc Aggregates
- Author
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Shin Chung Kang, Ruliang Li, Tao Pan, Thomas Wisniewski, Chaoyang Li, Po Tein, Geoff Barnard, Ian McConnell, John D. Robinson, David R. Brown, Man Sun Sy, Shaoman Yin, Poki Wong, Binggong Chang, and Andrew R. Thompsett
- Subjects
PrPSc Proteins ,Prions ,Protein Conformation ,medicine.drug_class ,animal diseases ,Immunology ,Enzyme-Linked Immunosorbent Assay ,Monoclonal antibody ,Sensitivity and Specificity ,Microbiology ,law.invention ,Mice ,Protein structure ,law ,Virology ,medicine ,Animals ,Prion protein ,Specific enzyme ,Endopeptidase K ,biology ,Antibodies, Monoclonal ,Brain ,Proteinase K ,Molecular biology ,Recombinant Proteins ,nervous system diseases ,Molecular Weight ,Insect Science ,biology.protein ,Recombinant DNA ,Pathogenesis and Immunity ,Dimerization - Abstract
The conversion of the normal cellular prion protein, PrP C , into the protease-resistant, scrapie PrP Sc aggregate is the cause of prion diseases. We developed a novel enzyme-linked immunosorbent assay (ELISA) that is specific for PrP aggregate by screening 30 anti-PrP monoclonal antibodies (MAbs) for their ability to react with recombinant mouse, ovine, bovine, or human PrP dimers. One MAb that reacts with all four recombinant PrP dimers also reacts with PrP Sc aggregates in ME7-, 139A-, or 22L-infected mouse brains. The PrP Sc aggregate is proteinase K resistant, has a mass of 2,000 kDa or more, and is present at a time when no protease-resistant PrP is detectable. This simple and sensitive assay provides the basis for the development of a diagnostic test for prion diseases in other species. Finally, the principle of the aggregate-specific ELISA we have developed may be applicable to other diseases caused by abnormal protein aggregation, such as Alzheimer's disease or Parkinson's disease.
- Published
- 2005