title: Sensitive detection of aggregated prion protein via proximity ligation.
abstract: The DNA assisted solid-phase proximity ligation assay (SP-PLA) provides a unique opportunity to specifically detect prion protein (PrP) aggregates by investigating the collocation of 3 or more copies of the specific protein. We have developed an SP-PLA that can detect PrP aggregates in brain homogenates from infected hamsters even after a 10(7)-fold dilution. In contrast, brain homogenate from uninfected animals did not generate a detectable signal at 100-fold higher concentration. Using either of the 2 monoclonal anti-PrP antibodies, 3F4 and 6H4, we successfully detected low concentrations of aggregated PrP. The presented results provide a proof of concept that this method might be an interesting tool in the development of diagnostic approaches of prion diseases.
authors.affiliation: a Department of Immunology, Genetics and Pathology; Science for Life Laboratory ; Uppsala University ; Uppsala , Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Hammond
authors.forename: Lotta
authors.initials: L
authors.lastname: Wik
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Deslys
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Comoy
authors.forename: Tommy
authors.initials: T
authors.lastname: Linné
authors.forename: Ulf
authors.initials: U
authors.lastname: Landegren
authors.forename: Masood
authors.initials: M
authors.lastname: Kamali-Moghaddam
mesh_terms: D000682:Amyloid / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D055910:Antibodies, Immobilized; D001923:Brain Chemistry; D006224:Cricetinae; D007118:Immunoassay / Q000379:methods*; D011328:Prions / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D000682:Amyloid; D055910:Antibodies, Immobilized; D011328:Prions
keywords: 263K; BSE, bovine spongiform encephalopathy; CJD, Creutzfeldt-Jakob disease; CSF, cerebrospinal fluid; FIDA, fluorescence intensity distribution analysis; PLA, proximity ligation assay; PMCA, protein misfolding cyclic amplification; PrP, prion protein; PrPC, cellular prion protein; PrPSc, scrapie prion protein; QuIC, quaking-induced conversion; SP-PLA, solid phase proximity ligation assay; diagnosis; monoclonal antibody; prion protein; proximity ligation assay; qPCR, quantitative real-time PCR