{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T11:08:01Z","timestamp":1789384081887,"version":"build-2803163510"},"reference-count":45,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100014188","name":"Korean government","doi-asserted-by":"publisher","award":["RS-2024-00440975"],"award-info":[{"award-number":["RS-2024-00440975"]}],"id":[{"id":"10.13039\/501100014188","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014188","name":"Korean government","doi-asserted-by":"publisher","award":["RS-2026-25476730"],"award-info":[{"award-number":["RS-2026-25476730"]}],"id":[{"id":"10.13039\/501100014188","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014188","name":"Korean government","doi-asserted-by":"publisher","award":["RS-2026-25510121"],"award-info":[{"award-number":["RS-2026-25510121"]}],"id":[{"id":"10.13039\/501100014188","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministry of Education and Busan Metropolitan City, Republic of Korea","award":["2026-glocal-02-004-511-004"],"award-info":[{"award-number":["2026-glocal-02-004-511-004"]}]},{"DOI":"10.13039\/501100014188","name":"Ministry of Science and ICT","doi-asserted-by":"publisher","award":["2710107319"],"award-info":[{"award-number":["2710107319"]}],"id":[{"id":"10.13039\/501100014188","id-type":"DOI","asserted-by":"publisher"}]},{"award":["2710107319"],"award-info":[{"award-number":["2710107319"]}],"id":[{"id":"https:\/\/ror.org\/01wpjm123","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Efficient evaluation of aptamer candidates after systematic evolution of ligands by exponential enrichment (SELEX) remains limited by the time and cost required for direct kinetic characterization. We developed a centrifugal microspin filtration (MSF)-based MSF\u2013koff assay for retention-based, dissociation-sensitive pre-screening of aptamer candidates. Repeated washing preferentially removes rapidly dissociating aptamers from target-immobilized magnetic beads, and the retained aptamers are quantified by quantitative polymerase chain reaction and expressed as Wash 0-normalized retention (W0-NRT). In a thrombin aptamer panel, W0-NRT showed a strong inverse relationship with SPR-derived koff. A preliminary analysis of streptavidin aptamers likewise showed a W0-NRT-based ranking consistent with the literature-reported koff values, although the two target systems exhibited distinct empirical relationships. In a wash-based thrombin sandwich assay, two affinity-matched sensor aptamers differing 1.76-fold in koff were ranked in the same order by post-wash retention as by W0-NRT. The agreement was in rank order rather than in the magnitude of retention. The parallel microspin filter format was also estimated to offer advantages in analysis time and assay-related cost. The MSF\u2013koff assay therefore provides a simple post-SELEX approach for prioritizing candidates with higher retention under dissociation-sensitive washing before detailed kinetic characterization.<\/jats:p>","DOI":"10.3390\/bios16090510","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T13:04:57Z","timestamp":1789045497000},"page":"510","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Retention-Based, Dissociation-Sensitive Pre-Screening of Aptamers Using a Centrifugal Microspin Filter"],"prefix":"10.3390","volume":"16","author":[{"given":"Cheeyoon","family":"Ahn","sequence":"first","affiliation":[{"name":"Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1832-4121","authenticated-orcid":false,"given":"Hwayeon","family":"Jeong","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Kyungjin","family":"Jeon","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"visualization","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Cheulhee","family":"Jung","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1038\/355850a0","article-title":"Selection in Vitro of Single-Stranded DNA Molecules That Fold into Specific Ligand-Binding Structures","volume":"355","author":"Ellington","year":"1992","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2540","DOI":"10.1021\/acssensors.4c00270","article-title":"Dual-Aptamer Recognition of DNA Logic Gate Sensor-Based Specific Exosomal Proteins for Ovarian Cancer Diagnosis","volume":"9","author":"Zhao","year":"2024","journal-title":"ACS Sens."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"116380","DOI":"10.1016\/j.bios.2024.116380","article-title":"CRISPR\/Cas12a and Aptamer-Chemiluminescence Based Analysis for the Relative Abundance Determination of Tumor-Related Protein Positive Exosomes for Breast Cancer Diagnosis","volume":"259","author":"Guan","year":"2024","journal-title":"Biosens. 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