Generation of Antibodies Against Prion Protein by Scrapie-Infected Cell Immunization of PrP ^0/0 Mice

Abstract

Four monoclonal antibodies (MAbs) specific for prion protein (PrP) were generated by using PrP-knockout mice immunized with a scrapie-infected mouse neuroblastoma cell line (N2a/22L). The MAbs reacted with both the cellular form (PrP(C)) and the protease K-treated form (PrP(Sc)) on Western blotting. Of the four MAbs, three recognized mouse and hamster PrP, while the remaining MAb recognized mouse, sheep, and bovine PrPs. In addition, these MAbs were shown to react only with the unglycosylated and monoglycoslated forms of PrP(Sc) in N2a/22L, but reacted with all glycosylated forms of PrP(C) and PrP(Sc) from mouse brain. This study was the first to report the development of anti-PrP MAbs using scrapie-infected cells as an immunogen and provides one approach for the generation of PrP-specific MAbs.