title: Guanidine hydrochloride extraction and detection of prion proteins in mouse and hamster prion diseases by ELISA.
abstract: Current detection of transmissible spongiform encephalopathy (TSE) relies on the proteolytic generation of a protease-resistant core from the scrapie isoform of prion protein (PrP(Sc)) followed by immunoblotting. This process is non-quantitative, time-consuming, and technically demanding. Recently, an alternative in vitro test for TSE based on the differential extraction of brain homogenates using guanidine hydrochloride followed by DELFIA (Dissociation Enhanced Lanthanide FluoroImmunoAssay) has been developed. In the present study, this approach was adopted using a panel of anti-PrP monoclonal antibodies (MAbs) in conventional sandwich enzyme-linked immunosorbent assay (ELISA) to investigate hamster and two distinct strains of mouse prion diseases. Although PrP species were present in both soluble and insoluble fractions from normal as well as TSE samples, only the PrP species in the insoluble fractions from the latter samples were protease-resistant. In addition, certain anti-PrP MAb pairs could distinguish the PrP species in infected brains from those in the normal samples. The ability to differentiate disease-associated PrP isoforms without proteinase K digestion could serve as a panacea for developing a reliable and rapid diagnostic test for prion diseases.
authors.affiliation: Institute of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA.
authors.forename: Shin-Chung
authors.initials: SC
authors.lastname: Kang
authors.forename: Ruliang
authors.initials: R
authors.lastname: Li
authors.forename: Chuanping
authors.initials: C
authors.lastname: Wang
authors.forename: Tao
authors.initials: T
authors.lastname: Pan
authors.forename: Tong
authors.initials: T
authors.lastname: Liu
authors.forename: Richard
authors.initials: R
authors.lastname: Rubenstein
authors.forename: Geoff
authors.initials: G
authors.lastname: Barnard
authors.forename: Boon-Seng
authors.initials: BS
authors.lastname: Wong
authors.forename: Man-Sun
authors.initials: MS
authors.lastname: Sy
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001923:Brain Chemistry; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D019791:Guanidine; D008647:Mesocricetus; D051379:Mice; D008815:Mice, Inbred Strains; D018620:PrPSc Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D017096:Prion Diseases / Q000175:diagnosis*; D011328:Prions / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D012995:Solubility; D013048:Specimen Handling / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D018620:PrPSc Proteins; D011328:Prions; D019791:Guanidine