title: Generation of monoclonal antibodies against human prion proteins in PrP0/0 mice.
abstract: BACKGROUND
  Prion diseases belong to a group of neurodegenerative disorders affecting humans and animals. The human diseases include kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome (GSS), and fatal familial insomnia (FFI). The pathogenic mechanisms of the prion diseases are not yet understood. Monoclonal antibodies provide valuable tools in the diagnosis, as well as in the basic research, of several diseases; however, monospecific antisera or monoclonal antibodies (mAbs) against human prion proteins were, until now, not available.
  
  
  METHODS
  We have developed an immunization protocol based on nucleic acid injection into nontolerant PrP0/0 mice. DNA or RNA coding for different human prion proteins including the mutated sequences associated with CJD, GSS, and FFI were injected into muscle tissue. Mice were primarily inoculated with DNA plasmids encoding the prion protein (PRNP) gene and boosted either with DNA, RNA, or recombinant Semliki Forest Virus particles expressing PRNP. Hybridomas were then prepared.
  
  
  RESULTS
  Different mAbs against human prion proteins were obtained, and their binding behavior was analyzed by peptide enzyme-linked immunosorbent assay, Western blot, immunofluorescence, and immunoprecipitation. Their cross-reactivity with prion protein from other species was also determined. Our mAbs are directed against four different linear epitopes and may also recognize discontinuous regions of the native prion protein.
  
  
  CONCLUSIONS
  These antibodies should allow us to address questions concerning the nature of the prion protein as well as the initiation and progression of prion diseases. Moreover, these mAbs can now be used for the diagnosis of prion diseases of humans and animals.
authors.affiliation: Department of Virology and Immunology, German Primate Center (DPZ), Goettingen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Krasemann
authors.forename: M H
authors.initials: MH
authors.lastname: Groschup
authors.forename: S
authors.initials: S
authors.lastname: Harmeyer
authors.forename: G
authors.initials: G
authors.lastname: Hunsmann
authors.forename: W
authors.initials: W
authors.lastname: Bodemer
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018604:Epitope Mapping; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007114:Immunization; D051379:Mice; D008815:Mice, Inbred Strains; D009154:Mutation / Q000235:genetics; D017096:Prion Diseases / Q000378:metabolism; D011328:Prions / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D012672:Semliki forest virus / Q000378:metabolism; D014162:Transfection / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D011328:Prions