{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,22]],"date-time":"2026-03-22T06:02:39Z","timestamp":1774159359342,"version":"3.50.1"},"reference-count":54,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,3,20]],"date-time":"2026-03-20T00:00:00Z","timestamp":1773964800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003478","name":"Ministry of Health, Labour and Welfare of Japan","doi-asserted-by":"publisher","award":["23KA1004"],"award-info":[{"award-number":["23KA1004"]}],"id":[{"id":"10.13039\/501100003478","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science, and Technology, Japan","doi-asserted-by":"crossref","award":["1801"],"award-info":[{"award-number":["1801"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001700","name":"Ministry of Education, Culture, Sports, Science, and Technology, Japan","doi-asserted-by":"crossref","award":["25K02167"],"award-info":[{"award-number":["25K02167"]}],"id":[{"id":"10.13039\/501100001700","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Real-time quaking-induced conversion (RT-QuIC) is highly sensitive for prion detection; however, inhibitory factors present in tissue homogenates readily interfere with the assay. We previously reported that recombinant cervid prion protein (rCerPrP) enabled the establishment of practical RT-QuIC for detecting chronic wasting disease and atypical bovine spongiform encephalopathy (BSE) prions, i.e., detecting low levels of prions in high concentration of brain tissue homogenates. Accordingly, the present study aimed to establish RT-QuIC for detecting classical BSE (C-BSE) and classical and atypical scrapie (C- and A-scrapie, respectively). A single-step lipid extraction using a 3:1 mixture of 2-butanol and methanol was effective as a pretreatment to remove inhibitors from brain homogenates. Among three rPrPs extensively evaluated, recombinant sheep PrP (rShPrP) was the most suitable substrate for practical detection of C-BSE prions. rCerPrP-173S\/177N and rCerPrP-98S\/173S\/177N, which carry sheep-type amino acid substations at codons 173 and 177 and at codons 98, 173, and 177, showed excellent performance for detecting C-scrapie prions. Moreover, rCerPrP-98S\/173S\/177N, but not rCerPrP-173S\/177N, was identified as an optimal substrate for detecting A-scrapie prions. These results suggested that combining inhibitor-removal pretreatment with the optimization of rPrP substrate for each animal prions further enhanced of RT-QuIC performance.<\/jats:p>","DOI":"10.3390\/pathogens15030333","type":"journal-article","created":{"date-parts":[[2026,3,20]],"date-time":"2026-03-20T11:44:46Z","timestamp":1774007086000},"page":"333","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Practical Real-Time Quaking-Induced Conversion for Detecting Classical Bovine Spongiform Encephalopathy and Classical and Atypical Scrapie Prions"],"prefix":"10.3390","volume":"15","author":[{"given":"Akio","family":"Suzuki","sequence":"first","affiliation":[{"name":"Laboratory of Veterinary Hygiene, Faculty of Veterinary Medicine, Graduate School of Infectious Diseases, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"},{"name":"One Health Research Center, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuhei","family":"Sawada","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Hygiene, Faculty of Veterinary Medicine, Graduate School of Infectious Diseases, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Taku","family":"Nakashima","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Hygiene, Faculty of Veterinary Medicine, Graduate School of Infectious Diseases, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toyotaka","family":"Sato","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Hygiene, Faculty of Veterinary Medicine, Graduate School of Infectious Diseases, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"},{"name":"One Health Research Center, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kohtaro","family":"Miyazawa","sequence":"additional","affiliation":[{"name":"Division of Infectious Animal Diseases, National Institute of Animal Health, Kannondai 3-1-5, Tsukuba 305-0856, Ibaraki, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3383-7918","authenticated-orcid":false,"given":"Yuichi","family":"Matsuura","sequence":"additional","affiliation":[{"name":"Division of Infectious Animal Diseases, National Institute of Animal Health, Kannondai 3-1-5, Tsukuba 305-0856, Ibaraki, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keigo","family":"Ikeda","sequence":"additional","affiliation":[{"name":"Division of Infectious Animal Diseases, National Institute of Animal Health, Kannondai 3-1-5, Tsukuba 305-0856, Ibaraki, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3459-7872","authenticated-orcid":false,"given":"Yoshifumi","family":"Iwamaru","sequence":"additional","affiliation":[{"name":"Division of Infectious Animal Diseases, National Institute of Animal Health, Kannondai 3-1-5, Tsukuba 305-0856, Ibaraki, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Motohiro","family":"Horiuchi","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Hygiene, Faculty of Veterinary Medicine, Graduate School of Infectious Diseases, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"},{"name":"One Health Research Center, Hokkaido University, Kita 18, Nishi 9, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"},{"name":"Veterinary Research Unit, International Institute for Zoonosis Control, Hokkaido University, Kita 20, Nishi 10, Kita-ku, Sapporo 060-0818, Hokkaido, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"13363","DOI":"10.1073\/pnas.95.23.13363","article-title":"Prions","volume":"95","author":"Prusiner","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1111\/bpa.12696","article-title":"Animal prion diseases: The risks to human health","volume":"29","author":"Houston","year":"2019","journal-title":"Brain Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3737","DOI":"10.1099\/vir.0.82011-0","article-title":"Infectious agent of sheep scrapie may persist in the environment for at least 16 years","volume":"87","author":"Georgsson","year":"2006","journal-title":"J. Gen. Virol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Mathiason, C.K., Hays, S.A., Powers, J., Hayes-Klug, J., Langenberg, J., Dahmes, S.J., Osborn, D.A., Miller, K.V., Warren, R.J., and Mason, G.L. (2009). Infectious prions in pre-clinical deer and transmission of chronic wasting disease solely by environmental exposure. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0005916"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1126\/science.1132661","article-title":"Infectious prions in the saliva and blood of deer with chronic wasting disease","volume":"314","author":"Mathiason","year":"2006","journal-title":"Science"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.3201\/eid1006.040010","article-title":"Environmental sources of prion transmission in mule deer","volume":"10","author":"Miller","year":"2004","journal-title":"Emerg. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1168","DOI":"10.1016\/j.celrep.2015.04.036","article-title":"Grass plants bind, retain, uptake, and transport infectious prions","volume":"11","author":"Pritzkow","year":"2015","journal-title":"Cell Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1016\/S0140-6736(96)91412-9","article-title":"A new variant of Creutzfeldt-Jakob disease in the UK","volume":"347","author":"Will","year":"1996","journal-title":"Lancet"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Baron, T., Vulin, J., Biacabe, A.G., Lakhdar, L., Verchere, J., Torres, J.M., and Bencsik, A. (2011). Emergence of classical BSE strain properties during serial passages of H-BSE in wild-type mice. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0015839"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Capobianco, R., Casalone, C., Suardi, S., Mangieri, M., Miccolo, C., Limido, L., Catania, M., Rossi, G., Di Fede, G., and Giaccone, G. (2007). Conversion of the BASE prion strain into the BSE strain: The origin of BSE?. PLoS Pathog., 3.","DOI":"10.1371\/journal.ppat.0030031"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"26853","DOI":"10.1073\/pnas.1915737116","article-title":"The emergence of classical BSE from atypical\/Nor98 scrapie","volume":"116","author":"Huor","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1186\/s13567-016-0394-1","article-title":"L-BSE experimentally transmitted to sheep presents as a unique disease phenotype","volume":"47","author":"Simmons","year":"2016","journal-title":"Vet. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1099\/vir.0.025387-0","article-title":"BSE infectivity in the absence of detectable PrPSc accumulation in the tongue and nasal mucosa of terminally diseased cattle","volume":"92","author":"Eiden","year":"2011","journal-title":"J. Gen. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.3201\/eid1607.091882","article-title":"Accumulation of L-type bovine prions in peripheral nerve tissues","volume":"16","author":"Iwamaru","year":"2010","journal-title":"Emerg. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1292\/jvms.19-0003","article-title":"Estimation of prion infectivity in tissues of cattle infected with atypical BSE by real time-quaking induced conversion assay","volume":"81","author":"Sawada","year":"2019","journal-title":"J. Vet. Med. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Suardi, S., Vimercati, C., Casalone, C., Gelmetti, D., Corona, C., Iulini, B., Mazza, M., Lombardi, G., Moda, F., and Ruggerone, M. (2012). Infectivity in skeletal muscle of cattle with atypical bovine spongiform encephalopathy. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0031449"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/nm.2294","article-title":"Ultrasensitive human prion detection in cerebrospinal fluid by real-time quaking-induced conversion","volume":"17","author":"Atarashi","year":"2011","journal-title":"Nat. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1099\/jgv.0.000367","article-title":"Sensitive and specific detection of classical scrapie prions in the brains of goats by real-time quaking-induced conversion","volume":"97","author":"Dassanayake","year":"2016","journal-title":"J. Gen. Virol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"McNulty, E., Nalls, A.V., Mellentine, S., Hughes, E., Pulscher, L., Hoover, E.A., and Mathiason, C.K. (2019). Comparison of conventional, amplification and bio-assay detection methods for a chronic wasting disease inoculum pool. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0216621"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e02162-16","DOI":"10.1128\/JVI.02162-16","article-title":"Endogenous Brain Lipids Inhibit Prion Amyloid Formation In Vitro","volume":"91","author":"Hoover","year":"2017","journal-title":"J. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e00078-11","DOI":"10.1128\/mBio.00078-11","article-title":"Prion disease blood test using immunoprecipitation and improved quaking-induced conversion","volume":"2","author":"Orru","year":"2011","journal-title":"mBio"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Wilham, J.M., Orru, C.D., Bessen, R.A., Atarashi, R., Sano, K., Race, B., Meade-White, K.D., Taubner, L.M., Timmes, A., and Caughey, B. (2010). Rapid end-point quantitation of prion seeding activity with sensitivity comparable to bioassays. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1001217"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2023","DOI":"10.1099\/jgv.0.000515","article-title":"Enhanced prion detection in biological samples by magnetic particle extraction and real-time quaking-induced conversion","volume":"97","author":"Denkers","year":"2016","journal-title":"J. Gen. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1099\/vir.0.069906-0","article-title":"Quantitative assessment of prion infectivity in tissues and body fluids by real-time quaking-induced conversion","volume":"96","author":"Henderson","year":"2015","journal-title":"J. Gen. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1080\/19336896.2020.1858694","article-title":"Involvement of N- and C-terminal region of recombinant cervid prion protein in its reactivity to CWD and atypical BSE prions in real-time quaking-induced conversion reaction in the presence of high concentrations of tissue homogenates","volume":"14","author":"Suzuki","year":"2020","journal-title":"Prion"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3852","DOI":"10.1128\/JVI.01740-08","article-title":"The region approximately between amino acids 81 and 137 of proteinase K-resistant PrPSc is critical for the infectivity of the Chandler prion strain","volume":"83","author":"Shindoh","year":"2009","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3421","DOI":"10.1128\/JCM.40.9.3421-3426.2002","article-title":"Biological and biochemical characterization of sheep scrapie in Japan","volume":"40","author":"Horiuchi","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1735","DOI":"10.1292\/jvms.21-0368","article-title":"Monitoring of chronic wasting disease using real-time quaking-induced conversion assay in Japan","volume":"83","author":"Suzuki","year":"2021","journal-title":"J. Vet. Med. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1042\/bj0820113","article-title":"Analysis of lipids from fresh and preserved adult human brains","volume":"82","author":"Davison","year":"1962","journal-title":"Biochem. J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1292\/jvms.66.515","article-title":"Effect of tissue deterioration on postmortem BSE diagnosis by immunobiochemical detection of an abnormal isoform of prion protein","volume":"66","author":"Hayashi","year":"2004","journal-title":"J. Vet. Med. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5766","DOI":"10.1021\/ac500317c","article-title":"Objective set of criteria for optimization of sample preparation procedures for ultra-high throughput untargeted blood plasma lipid profiling by ultra performance liquid chromatography-mass spectrometry","volume":"86","author":"Sarafian","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.jchromb.2017.08.020","article-title":"One-step lipid extraction for plasma lipidomics analysis by liquid chromatography mass spectrometry","volume":"1063","author":"Satomi","year":"2017","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1690","DOI":"10.1194\/jlr.D023036","article-title":"The BUME method: A novel automated chloroform-free 96-well total lipid extraction method for blood plasma","volume":"53","author":"Lofgren","year":"2012","journal-title":"J. Lipid Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5287","DOI":"10.1007\/s12035-018-1443-8","article-title":"Insights into the Bidirectional Properties of the Sheep-Deer Prion Transmission Barrier","volume":"56","author":"Harrathi","year":"2019","journal-title":"Mol. Neurobiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/S0021-9258(18)64849-5","article-title":"A simple method for the isolation and purification of total lipides from animal tissues","volume":"226","author":"Folch","year":"1957","journal-title":"J. Biol. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2667","DOI":"10.1096\/fj.13-246777","article-title":"Heparin binding confers prion stability and impairs its aggregation","volume":"28","author":"Vieira","year":"2014","journal-title":"FASEB J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4074","DOI":"10.1021\/acs.nanolett.3c01001","article-title":"Nanoparticle-Enhanced RT-QuIC (Nano-QuIC) Diagnostic Assay for Misfolded Proteins","volume":"23","author":"Christenson","year":"2023","journal-title":"Nano Lett."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Hwang, S., Dassanayake, R.P., and Nicholson, E.M. (2019). PAD-Beads enrichment enhances detection of PrP(Sc) using real-time quaking-induced conversion. BMC Res. Notes, 12.","DOI":"10.1186\/s13104-019-4842-7"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Thomas, C.M., Salamat, M.K.F., de Wolf, C., McCutcheon, S., Blanco, A.R.A., Manson, J.C., Hunter, N., and Houston, E.F. (2023). Development of a sensitive real-time quaking-induced conversion (RT-QuIC) assay for application in prion-infected blood. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0293845"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Hwang, S., Greenlee, J.J., and Nicholson, E.M. (2017). Use of bovine recombinant prion protein and real-time quaking-induced conversion to detect cattle transmissible mink encephalopathy prions and discriminate classical and atypical L- and H-Type bovine spongiform encephalopathy. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0172391"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Levavasseur, E., Biacabe, A.G., Comoy, E., Culeux, A., Grznarova, K., Privat, N., Simoneau, S., Flan, B., Sazdovitch, V., and Seilhean, D. (2017). Detection and partial discrimination of atypical and classical bovine spongiform encephalopathies in cattle and primates using real-time quaking-induced conversion assay. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0172428"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1128\/JCM.02731-15","article-title":"Detection of Atypical H-Type Bovine Spongiform Encephalopathy and Discrimination of Bovine Prion Strains by Real-Time Quaking-Induced Conversion","volume":"54","author":"Masujin","year":"2016","journal-title":"J. Clin. Microbiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1128\/JCM.02906-14","article-title":"Detection and discrimination of classical and atypical L-type bovine spongiform encephalopathy by real-time quaking-induced conversion","volume":"53","author":"Orru","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Orru, C.D., Groveman, B.R., Raymond, L.D., Hughson, A.G., Nonno, R., Zou, W., Ghetti, B., Gambetti, P., and Caughey, B. (2015). Bank Vole Prion Protein As an Apparently Universal Substrate for RT-QuIC-Based Detection and Discrimination of Prion Strains. PLoS Pathog., 11.","DOI":"10.1371\/journal.ppat.1004983"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1049","DOI":"10.1016\/j.bbrc.2020.03.183","article-title":"Discrimination between L-type and C-type bovine spongiform encephalopathy by the strain-specific reactions of real-time quaking-induced conversion","volume":"526","author":"Ubagai","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"6173","DOI":"10.1038\/s41598-019-42449-7","article-title":"Early and Pre-Clinical Detection of Prion Seeding Activity in Cerebrospinal Fluid of Goats using Real-Time Quaking-Induced Conversion Assay","volume":"9","author":"Favole","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"102381","DOI":"10.1016\/j.jbc.2022.102381","article-title":"Central residues in prion protein PrP(C) are crucial for its conversion into the pathogenic isoform","volume":"298","author":"Pasiana","year":"2022","journal-title":"J. Biol. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Uchiyama, K., Miyata, H., Yamaguchi, Y., Imamura, M., Okazaki, M., Pasiana, A.D., Chida, J., Hara, H., Atarashi, R., and Watanabe, H. (2020). Strain-Dependent Prion Infection in Mice Expressing Prion Protein with Deletion of Central Residues 91\u2013106. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21197260"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"106620","DOI":"10.1016\/j.bpc.2021.106620","article-title":"Free energy simulations to understand the effect of Met \u2192 Ala mutations at positions 205, 206 and 213 on stability of human prion protein","volume":"275","author":"Lee","year":"2021","journal-title":"Biophys. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"28263","DOI":"10.1074\/jbc.M112.354779","article-title":"Methionine oxidation perturbs the structural core of the prion protein and suggests a generic misfolding pathway","volume":"287","author":"Younan","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"19076","DOI":"10.1074\/jbc.M117.794107","article-title":"Asparagine and glutamine ladders promote cross-species prion conversion","volume":"292","author":"Kurt","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"937","DOI":"10.1099\/0022-1317-79-4-937","article-title":"Synthetic peptide vaccines yield monoclonal antibodies to cellular and pathological prion proteins of ruminants","volume":"79","author":"Harmeyer","year":"1998","journal-title":"J. Gen. Virol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1111\/j.1348-0421.2009.00190.x","article-title":"A novel anti-prion protein monoclonal antibody and its single-chain fragment variable derivative with ability to inhibit abnormal prion protein accumulation in cultured cells","volume":"54","author":"Shimizu","year":"2010","journal-title":"Microbiol. Immunol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1111\/j.1348-0421.2007.tb03954.x","article-title":"Characterization of prion susceptibility in Neuro2a mouse neuroblastoma cell subclones","volume":"51","author":"Uryu","year":"2007","journal-title":"Microbiol. Immunol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/3\/333\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,22]],"date-time":"2026-03-22T05:28:21Z","timestamp":1774157301000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/3\/333"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,20]]},"references-count":54,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["pathogens15030333"],"URL":"https:\/\/doi.org\/10.3390\/pathogens15030333","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,20]]}}}