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Here, based on de novo protein fragmentation, the EASY MHC-I (EZ MHC-I) assay favors peptide antigen screening to an unheralded hands-on time of seconds per peptide due to the empty single chain MHC-I protein instability. Unlike tedious traditional labeling- and antibody-based MHC-I assays, repurposed enterokinase directly fragments the unstable single MHC-I chain protein unless rescued by a stabilizing peptide under luminal condition. Herein, the principle behind EZ MHC-I assay not only characterizes the overlooked stability as a known better indicator of immunogenicity than classical affinity but also the novel use of enterokinase from the duodenum to target destabilized MHC-I protein not bearing the standard Asp-Asp-Asp-Asp-Lys motif, which may protend to other protein instability-based assays.<\/jats:p>","DOI":"10.1038\/s41598-020-71785-2","type":"journal-article","created":{"date-parts":[[2020,9,10]],"date-time":"2020-09-10T10:03:14Z","timestamp":1599732194000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Destabilizing single chain major histocompatibility complex class I protein for repurposed enterokinase proteolysis"],"prefix":"10.1038","volume":"10","author":[{"given":"Jackwee","family":"Lim","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2020,9,10]]},"reference":[{"key":"71785_CR1","doi-asserted-by":"publisher","first-page":"382","DOI":"10.1038\/ni1175","volume":"6","author":"SJ Turner","year":"2005","unstructured":"Turner, S. 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