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Herein, we simultaneously identified the mutant A21G, E22G, E22Q, and the wild\u2010type (WT) A\u03b2<jats:sub>18\u201326<\/jats:sub> peptides with aerolysin nanopore using a 3D blockage mapping strategy. The standard deviation of current blockade fluctuations (<jats:italic>\u03c3<\/jats:italic><jats:sub>b<\/jats:sub>) was proposed as a new supplement to current blockage (<jats:italic>I<\/jats:italic><jats:sub>b<\/jats:sub>\/<jats:italic>I<\/jats:italic><jats:sub>0<\/jats:sub>) and duration time (<jats:italic>t<\/jats:italic><jats:sub>D<\/jats:sub>) to profile the blockage characteristics of single molecules. Although the WT and A21G A\u03b2<jats:sub>18\u201026<\/jats:sub> are indistinguishable in a traditional <jats:italic>I<\/jats:italic><jats:sub>b<\/jats:sub>\/<jats:italic>I<\/jats:italic><jats:sub>0<\/jats:sub>\u2010<jats:italic>t<\/jats:italic><jats:sub>D<\/jats:sub> 2D description, \u223c87\u2009% of the blockade events can be accurately classified with half reduction of false identification using a combination of <jats:italic>I<\/jats:italic><jats:sub>b<\/jats:sub>\/<jats:italic>I<\/jats:italic><jats:sub>0<\/jats:sub>, <jats:italic>t<\/jats:italic><jats:sub>D,<\/jats:sub> and <jats:italic>\u03c3<\/jats:italic><jats:sub>b<\/jats:sub>. This work offers an easy and reliable strategy to promote nanopore sensitivity of peptide mutants, leading to a more precise analysis of pathogenic mutations for developing effective diagnosis and treatment.<\/jats:p>","DOI":"10.1002\/anie.202209970","type":"journal-article","created":{"date-parts":[[2022,9,9]],"date-time":"2022-09-09T21:22:12Z","timestamp":1662758532000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":45,"title":["3D Blockage Mapping for Identifying Familial Point Mutations in Single Amyloid\u2010\u03b2 Peptides with a Nanopore"],"prefix":"10.1002","volume":"61","author":[{"given":"Kai\u2010Li","family":"Xin","sequence":"first","affiliation":[{"name":"State Key Laboratory of Analytical Chemistry for Life Science School of Chemistry and Chemical Engineering Nanjing University  Nanjing 210023 P. R. 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