{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,26]],"date-time":"2026-10-26T18:40:12Z","timestamp":1793040012574,"version":"4.5.0"},"reference-count":17,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T00:00:00Z","timestamp":1791763200000},"content-version":"am","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Lampe Joint Department of Biomedical Engineering","id":[{"id":"https:\/\/ror.org\/03hakdk41","id-type":"ROR","asserted-by":"crossref"}]},{"DOI":"10.13039\/100007028","name":"Leona M. and Harry B. Helmsley Charitable Trust","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007028","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>To realize the continuous monitoring of insulin, the development of a regenerable engineered antibody is expected, whose binding constant is modulated by mild in situ stimulation for future in vivo application. In this study, we aimed to develop pH-responsive anti-insulin antibodies based on a clinically established analytical antibody, the HUI-018 anti-insulin antibody. Its high-resolution co-crystal structure with human insulin (Protein Data Bank (PDB) ID: 6Z7W) provides a uniquely precise blueprint for molecular engineering by preparing engineered HUI-018 single-chain variable fragment. Among the engineered variants, the N93H\/E101K mutant exhibited the greatest pH-responsiveness, showing a 5.2-fold increased dissociation rate at pH\u00a06.0 while maintaining binding affinity and specificity comparable to wild-type (WT) at pH\u00a07.4. Repeated cycles of insulin binding at pH\u00a07.4 and regeneration at pH\u00a06.0 demonstrated that the N93H\/E101K mutant achieved substantially higher regeneration and rebinding efficiencies than WT. These results demonstrate that an engineered pH-responsive antibody enables efficient binding site regeneration through a mild pH change. To our knowledge, this is the first report of an anti-insulin antibody capable of binding site regeneration at pH\u00a06.0, representing an important step toward antibody-based continuous insulin monitoring and continuous sensing of protein biomarkers.<\/jats:p>","DOI":"10.1093\/jb\/mvag071","type":"journal-article","created":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T11:48:30Z","timestamp":1791546510000},"source":"Crossref","is-referenced-by-count":0,"title":["Engineering of pH-regenerable anti-insulin single chain variable fragments for continuous insulin monitoring"],"prefix":"10.1093","author":[{"given":"Qianming","family":"Xu","sequence":"first","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ellie D","family":"Wilson","sequence":"additional","affiliation":[{"name":"Lampe Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill , 10202 Mary Ellen Jones Building, UNC Chapel Hill, NC 27599 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mai","family":"Hamasaki","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hirobumi","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayumi","family":"Tanaka","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junko","family":"Okuda-Shimazaki","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miho","family":"Oda","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daimei","family":"Miura","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koji","family":"Sode","sequence":"additional","affiliation":[{"name":"Lampe Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill , 10202 Mary Ellen Jones Building, UNC Chapel Hill, NC 27599 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6795-8377","authenticated-orcid":false,"given":"Ryutaro","family":"Asano","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology , 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,10,12]]},"reference":[{"key":"2026102614212833000_ref1","doi-asserted-by":"crossref","DOI":"10.1016\/j.bios.2021.113054","article-title":"Continuous glucose monitoring systems-current status and future perspectives of the flagship technologies in biosensor research","volume":"181","author":"Lee","year":"2021","journal-title":"Biosens. 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