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Journal of Virology, October 2005, p. 12355-12364, Vol. 79, No. 19
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.19.12355-12364.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

An Aggregation-Specific Enzyme-Linked Immunosorbent Assay: Detection of Conformational Differences between Recombinant PrP Protein Dimers and PrP^Sc Aggregates

Tao Pan,^1, Binggong Chang,¹ Poki Wong,¹ Chaoyang Li,¹ Ruliang Li,¹ Shin-Chung Kang,¹ John D. Robinson,² Andrew R. Thompsett,³ Po Tein,⁵ Shaoman Yin,⁵ Geoff Barnard,⁶ Ian McConnell,⁶ David R. Brown,³ Thomas Wisniewski,⁴ and Man-Sun Sy^2*

Institute of Pathology,¹ Department of Neurosciences, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44107-1712,² Department of Biology and Biochemistry, Bath University, Bath, United Kingdom,³ Department of Neurology, Psychiatry and Pathology, New York University School of Medicine, New York, New York 10016,⁴ Institute of Microbiology, Chinese Academy of Science, Beijing 100080, People's Republic of China,⁵ Centre for Veterinary Science, Department of Clinical Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom⁶

Received 22 April 2005/ Accepted 1 July 2005

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.

Present address: Adlyfe Inc., 9430 Key West Ave., Rockville, MD 20850.

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