Sensitive detection of aggregated prion protein via proximity ligation |
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| Authors: | Maria Hammond Lotta Wik Jean-Philippe Deslys Emmanuel Comoy Tommy Linné Ulf Landegren Masood Kamali-Moghaddam |
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| Affiliation: | 1Department of Immunology, Genetics and Pathology; Science for Life Laboratory; Uppsala University; Uppsala, Sweden;2Institute of Emerging Diseases and Innovative Therapies (iMETI); Division of Prions and Related Diseases (SEPIA); Atomic Energy Commission (CEA); Fontenay-aux-Roses, France;3Department of Biomedical Sciences and Veterinary Public Health; Swedish University of Agricultural Sciences; Uppsala, Sweden |
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| 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 107-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. |
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| Keywords: | diagnosis monoclonal antibody prion protein proximity ligation assay 263K |
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