{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:35:23Z","timestamp":1787027723624,"version":"build-2736575974"},"reference-count":66,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T00:00:00Z","timestamp":1762300800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection","award":["FHPB2023-3"],"award-info":[{"award-number":["FHPB2023-3"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Over the past few decades, Fc fragment-conjugated proteins, such as Protein A, have been extensively utilized across a range of applications, including antibody purification, site-specific immobilization of antibodies, and the development of biosensing platforms. In this study, building upon our group prior research, we designed and screened an affinity DNA functional ligand (A-DNAFL) and experimentally validated its binding affinity (KD = 6.59 \u00d7 10\u22128) toward mouse IgG antibodies, whose binding performance was comparable to that of protein A. Systematic evaluations were performed to assess the binding efficiency under varying pH levels and ionic strength conditions. Optimal antibody immobilization was achieved in PBST-B buffer under physiological pH 7.2\u20137.4 and containing approximately 154 mM Na+ and 4 mM K+. Two competitive binding assays confirmed that the A-DNAFL binds to the Fc fragment of murine IgG antibody. Furthermore, molecular docking simulations were employed to investigate the interaction mode, revealing key residues involved in binding as well as the contributions of hydrogen bonding and hydrophobic interactions to complex stabilization. Leveraging these insights, A-DNAFL was utilized as a tool for oriented immobilization of antibodies on the sensing interface, enabling the construction of an immunosensor for ricin detection. Following optimization of immobilization parameters, the biosensor exhibited a detection limit of 30.5 ng\/mL with the linear regression equation is lg(Response) = 0.329 lg(Cricin) \u2212 2.027 (N = 9, R = 0.938, p &lt; 0.001)\u2014representing a 64-fold improvement compared to conventional protein A-based methods. The system demonstrated robust resistance to nonspecific interference. Sensing interface reusability was also evaluated, showing only 8.55% signal reduction after two regeneration cycles, indicating that glycine effectively elutes bound antibodies while preserving sensor activity. In summary, the A-DNAFL presented in this study represents a novel antibody-directed immobilization material that serves as a promising alternative to protein A. It offers several advantages, including high modifiability, low production cost, and a relatively small molecular weight. These features collectively contribute to its broad application potential in biosensing, antibody purification, and other areas of life science research.<\/jats:p>","DOI":"10.3390\/bios15110747","type":"journal-article","created":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T17:06:06Z","timestamp":1762362366000},"page":"747","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Construction of an Immunosensor Based on the Affinity DNA Functional Ligands to the Fc Segment of IgG Antibody"],"prefix":"10.3390","volume":"15","author":[{"given":"Qianyu","family":"Yang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiwei","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinrui","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8808-8235","authenticated-orcid":false,"given":"Zihao","family":"Zhao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ze","family":"Fan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Du","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianjie","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2423-8123","authenticated-orcid":false,"given":"Jiwei","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiang","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xihui","family":"Mu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaoyang","family":"Tong","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Chemistry for NBC Hazards Protection, Beijing 102205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.xphs.2019.05.031","article-title":"Considerations for the Design of Antibody-Based Therapeutics","volume":"109","author":"Goulet","year":"2020","journal-title":"J. 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