title: Production and characterization of a panel of monoclonal antibodies against native human cellular prion protein.
abstract: The human prion diseases, such as variant Creutzfeldt-Jakob disease (vCJD), are characterized by the conversion of the normal cellular prion protein (PrP(C)) into an abnormal disease associated form (PrP(Sc)). Monoclonal antibodies (MAbs) that recognize these different PrP isoforms are valuable reagents both in the diagnosis of these diseases and in prion disease research in general but we know of no attempts to raise MAbs against native human PrP(C). We immunized prion protein gene ablated (PrP(-/-)) mice with native human PrP(C) purified from platelets (pHuPrP) generating a predominantly IgG isotype anti-pHuPrP polyclonal antibody response in all mice. Following fusion of splenocytes from the immunized mice with SP2/0 myeloma cells, we were able to identify single cell clone and cryopreserve 14 stable hybridoma cell lines producing MAbs that reacted with pHuPrP. The properties of these MAbs (such as isotype, binding to native/denatured pHuPrP, and HuPrP epitopes recognized) are described. Furthermore, several of these MAbs showed a selectivity in their ability to immunoprecipitate disease associated PrP(Sc) and its corresponding protease resistant core (PrP(res)).
authors.affiliation: National CJD Surveillance Unit, School of Molecular and Clinical Medicine (Pathology), University of Edinburgh, Western General Hospital, Edinburgh EH4 2XU, United Kingdom.
authors.forename: Michael
authors.initials: M
authors.lastname: Jones
authors.forename: Victoria
authors.initials: V
authors.lastname: McLoughlin
authors.forename: John G
authors.initials: JG
authors.lastname: Connolly
authors.forename: Christine F
authors.initials: CF
authors.lastname: Farquhar
authors.forename: Ian R
authors.initials: IR
authors.lastname: MacGregor
authors.forename: Mark W
authors.initials: MW
authors.lastname: Head
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D007562:Creutzfeldt-Jakob Syndrome / Q000175:diagnosis / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D006801:Humans; D006825:Hybridomas; D047468:Immunoprecipitation; D051379:Mice; D018345:Mice, Knockout; D018621:PrPC Proteins / Q000276:immunology* / Q000302:isolation & purification; D018620:PrPSc Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D018620:PrPSc Proteins