BioSpace Collaborative - Influence of Mabs on PrPSc Formation Using In Vitro and Cell-Free Systems

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Neurological Disorders - Neuroscience - Immunology - Infectious Diseases

Influence of Mabs on PrPSc Formation Using In Vitro and Cell-Free Systems
Published: Friday, July 27, 2012
Author: Binggong Chang et al.

by Binggong Chang, Robert Petersen, Thomas Wisniewski, Richard Rubenstein

PrP^Sc is believed to serve as a template for the conversion of PrP^C to the abnormal isoform. This process requires contact between the two proteins and implies that there may be critical contact sites that are important for conversion. We hypothesized that antibodies binding to either PrP^cor PrP^Sc would hinder or prevent the formation of the PrP^C–PrP^Sc complex and thus slow down or prevent the conversion process. Two systems were used to analyze the effect of different antibodies on PrP^Sc formation: (i) neuroblastoma cells persistently infected with the 22L mouse-adapted scrapie stain, and (ii) protein misfolding cyclic amplification (PMCA), which uses PrP^Sc as a template or seed, and a series of incubations and sonications, to convert PrP^C to PrP^Sc. The two systems yielded similar results, in most cases, and demonstrate that PrP-specific monoclonal antibodies (Mabs) vary in their ability to inhibit the PrP^C–PrP^Sc conversion process. Based on the numerous and varied Mabs analyzed, the inhibitory effect does not appear to be epitope specific, related to PrP^C conformation, or to cell membrane localization, but is influenced by the targeted PrP region (amino vs carboxy). More...