Inactivation of Prions and Amyloid Seeds with Hypochlorous Acid
Synthetic tau seeds were generated with recombinant K19 Cys-free tau fragment and treated with or without a 100-fold excess of BrioHOCl. The seed preparations were subjected to SDS-PAGE with non-specific staining for protein (Deep Purple) (A), immunoblotting probed with anti-tau antibody (B) or a seeded polymerization assay (C). In the seeded polymerization assay, the designated dilutions of the untreated (red) or BrioHOCl-treated (blue) seed samples were tested. To control for potential effects of BrioHOCl on the assay itself without allowing time for prior interactions the amyloid seed on its own, an amount of BrioHOCl comparable to that in a 10−3 dilution of treated seed was added directly to reactions solutions that were either left unseeded (black) or seeded with a 10−3 dilution of untreated seed (green). Reactions mock-seeded with 10−5 dilutions of NBH samples that were treated or not with 20 or 100 volumes of BrioHOCl are all shown in gray. Similar effects on K19 Cys-free seeding activity were obtained in at least five independent experiments.