Heterogeneity of Abnormal Prion Protein (PrP^Sc) in Murine Scrapie Prions Determined by PrP^Sc-Specific Monoclonal Antibodies

Abstract

In prion diseases, abnormal prion protein (PrPSc) is considered as the main component of the infectious agent. Delineation of PrPSc conformation is expected to be a critical factor in understanding properties of prions. However, practical methods to differentiate between conformers of PrPSc are inadequate. Here, we used two PrPSc-specific monoclonal antibodies (mAbs), 3B7 and 3H6, and found that mAb 3H6 detected a limited portion of PrPSc in five mice-adapted prion strains. The quantity of mAb 3H6-precipitated PrPSc was significantly lesser in 22L compared to other strains. This result provides a direct evidence of the conformational heterogeneity of PrPSc within the prion strains. Conformation-specific probes, like these mAbs, have the potential to be powerful tools for investigating conformational variations in PrPSc.

Selective propagation of mouse-passaged scrapie prions with long incubation period from a mixed prion population using GT1-7 cells

“…Protein G Dynabeads (Life Technologies Japan, Tokyo, Japan) were blocked with 4% (w/v) blocking solution (BlockAce; DS Pharma Biomedical, Osaka, Japan). Diluted supernatants (250 μL) were incubated with PrP Sc -specific monoclonal antibody (mAb) 3H6 [ 12 , 13 ] at a concentration of 1 μg/mL in 500-μL reaction volumes for 2.5 h with rotation at room temperature. Protein G Dynabeads (40 μL) were added and incubated for 2 h on the rotor at room temperature.…”