{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T05:12:56Z","timestamp":1767157976863,"version":"3.48.0"},"reference-count":48,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T00:00:00Z","timestamp":1766534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["5R21HD109743-02A1"],"award-info":[{"award-number":["5R21HD109743-02A1"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["1R03AG070511-01A1"],"award-info":[{"award-number":["1R03AG070511-01A1"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Metalloproteinases (MPs) are zinc-dependent endopeptidases, including matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinases (ADAMs), implicated in various diseases such as cancer, neurodegenerative disorders, and cardiovascular conditions. Among MPs, ADAM-17, also known as tumor necrosis factor-\u03b1 (TNF-\u03b1)-converting enzyme (TACE), plays a crucial role in extracellular matrix remodeling and cytokine release. Dysregulation of ADAM-17 contributes to inflammatory diseases, cancer progression, and immune modulation. While small-molecule inhibitors have been limited by off-target effects and instability, antibody-based approaches offer a more selective strategy. Monoclonal antibodies show promise in blocking ADAM-17 activity, but there are concerns about toxicity due to the lack of selectivity. Enhancing the binding affinity and selectivity of single-chain antibodies requires unraveling the structural details that drive MP targeting. This study uses yeast surface display (YSD) and fluorescence-activated cell sorting (FACS) to engineer single-chain variable fragment (scFv) antibodies with optimized complementarity-determining region 3 of the heavy chain (CDR-H3) conformations. Next-generation sequencing (NGS) was used to identify key residues contributing to high-affinity ADAM-17 binding. These findings offer a framework for designing monoclonal antibodies against ADAM-17 and other MPs, paving the way for novel antibody-based designer scaffolds with applications in developing therapeutics.<\/jats:p>","DOI":"10.3390\/biom16010031","type":"journal-article","created":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T14:27:51Z","timestamp":1766586471000},"page":"31","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Engineering Single-Chain Antibody Fragment (scFv) Variants Targeting A Disintegrin and Metalloproteinase-17 (ADAM-17)"],"prefix":"10.3390","volume":"16","author":[{"given":"Masoud","family":"Kalantar","sequence":"first","affiliation":[{"name":"Department of Chemical and Materials Engineering, University of Nevada, Reno, NV 89557, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7774-4604","authenticated-orcid":false,"given":"Elham","family":"Khorasani Buxton","sequence":"additional","affiliation":[{"name":"Department of Computer Science, University of Illinois, Springfield, IL 62703, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9077-3266","authenticated-orcid":false,"given":"Korey M.","family":"Reid","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Nevada, Reno, NV 89557, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Donald","family":"Bleyl","sequence":"additional","affiliation":[{"name":"Department of Computer Science, University of Illinois, Springfield, IL 62703, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3105-818X","authenticated-orcid":false,"given":"David M.","family":"Leitner","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Nevada, Reno, NV 89557, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2946-5584","authenticated-orcid":false,"given":"Maryam","family":"Raeeszadeh-Sarmazdeh","sequence":"additional","affiliation":[{"name":"Department of Chemical and Materials Engineering, University of Nevada, Reno, NV 89557, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Sampaio Moura, N., Schledwitz, A., Alizadeh, M., Patil, S.A., and Raufman, J.P. 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