An aggregation-specific enzyme-linked immunosorbent assay: detection of conformational differences between recombinant PrP protein dimers and PrP(Sc) aggregates.
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.
Pan
Tao
T
Institute of Pathology, School of Medicine, Case Western Reserve University, Cleveland, OH 44107-1712, USA.
Chang
Binggong
B
Wong
Poki
P
Li
Chaoyang
C
Li
Ruliang
R
Kang
Shin-Chung
SC
Robinson
John D
JD
Thompsett
Andrew R
AR
Tein
Po
P
Yin
Shaoman
S
Barnard
Geoff
G
McConnell
Ian
I
Brown
David R
DR
Wisniewski
Thomas
T
Sy
Man-Sun
MS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001921:Brain / Q000473:pathology; D019281:Dimerization; D019286:Endopeptidase K / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D051379:Mice; D008970:Molecular Weight; D018620:PrPSc Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D011328:Prions / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D018620:PrPSc Proteins; D011328:Prions; D011994:Recombinant Proteins; D019286:Endopeptidase K