{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T18:07:33Z","timestamp":1780078053802,"version":"3.54.0"},"reference-count":22,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T00:00:00Z","timestamp":1780012800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation of Shandong Province of China","award":["ZR2023MH021"],"award-info":[{"award-number":["ZR2023MH021"]}]},{"name":"National Natural Science Foundation of Shandong Province of China","award":["ZR2024QC347"],"award-info":[{"award-number":["ZR2024QC347"]}]},{"name":"Special Program for Enhancing Clinical Research Capacity at University of Health and Rehabilitation Sciences","award":["K2025LC7408"],"award-info":[{"award-number":["K2025LC7408"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibodies"],"abstract":"<jats:p>Background: The monkeypox virus (MPXV) has attracted considerable global attention due to its potential to cause widespread outbreaks, necessitating the development of rapid and accurate diagnostic methods of significant clinical importance. A29, a key envelope protein of MPXV, represents a promising diagnostic target. Methods: A novel monoclonal antibody, D10, was isolated from the human Tomlinson I+J phage display library by biopanning against the recombinant A29 protein. The D10 Fab fragment was expressed and purified, and its binding affinity was characterized by biolayer interferometry. Molecular docking was performed to predict potential interacting residues. Specificity and detection performance were evaluated by direct and competitive enzyme-linked immunosorbent assay (ELISA). Results: D10 possesses a unique complementarity-determining region sequence and exhibits strong binding affinity toward the A29 protein. Structural modeling analysis suggested potential interacting residues of A29, including Gln67, Arg74, Asn75, Arg81, and Asn84, which may primarily interact with Ser10, Thr5, Gly49, Gly47, and Glu97 in the heavy chain of D10. The binding affinity, determined by biolayer interferometry, showed a dissociation equilibrium constant of 6.44 nM, indicating strong binding capability. Furthermore, competitive ELISA demonstrated that D10 binds selectively to the A29 protein, with a half-maximal inhibitory concentration of 1.88 \u03bcg\/mL and a limit of detection of 0.12 \u03bcg\/mL. Conclusions: Overall, this monoclonal antibody provides a valuable tool for the immunological detection of MPXV and holds potential for future clinical diagnostic applications.<\/jats:p>","DOI":"10.3390\/antib15030045","type":"journal-article","created":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T17:36:18Z","timestamp":1780076178000},"page":"45","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Novel Monoclonal Antibody Targeting the A29 Protein of Monkeypox Virus and Its Application in Immunoassay"],"prefix":"10.3390","volume":"15","author":[{"given":"Nan","family":"Jia","sequence":"first","affiliation":[{"name":"School of Life Science and Technology, Shandong Second Medical University, Weifang 261053, China"},{"name":"School of Rehabilitation Sciences and Engineering, University of Health and Rehabilitation Sciences, Qingdao 266113, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weixiao","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Life Science and Technology, Shandong Second Medical University, Weifang 261053, China"},{"name":"School of Rehabilitation Sciences and Engineering, University of Health and Rehabilitation Sciences, Qingdao 266113, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangwei","family":"Zhao","sequence":"additional","affiliation":[{"name":"School of Rehabilitation Sciences and Engineering, University of Health and Rehabilitation Sciences, Qingdao 266113, China"},{"name":"School of Basic Medical Sciences, Shandong University, Ji\u2019nan 250014, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danfei","family":"Meng","sequence":"additional","affiliation":[{"name":"School of Rehabilitation Sciences and Engineering, University of Health and Rehabilitation Sciences, Qingdao 266113, China"},{"name":"School of Basic Medicine, Qingdao University, Qingdao 266071, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liyuan","family":"Zheng","sequence":"additional","affiliation":[{"name":"School of Rehabilitation Sciences and Engineering, University of Health and Rehabilitation Sciences, Qingdao 266113, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7417-1932","authenticated-orcid":false,"given":"Jinhua","family":"Dong","sequence":"additional","affiliation":[{"name":"School of Rehabilitation Sciences and Engineering, University of Health and Rehabilitation Sciences, Qingdao 266113, China"},{"name":"Institute of Integrated Research, Institute of Science Tokyo, Yokohama 226-8503, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.imj.2023.11.001","article-title":"Monkeypox virus (MPXV): A Brief account of global spread, epidemiology, virology, clinical features, pathogenesis, and therapeutic interventions","volume":"2","author":"Islam","year":"2023","journal-title":"Infect. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"76","DOI":"10.2174\/0118715265258451231214063506","article-title":"Monkeypox Virus (MPXV) Infection: A Review","volume":"24","author":"Kumari","year":"2024","journal-title":"Infect. Disord. Drug Targets"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Khan, A., Erickson, T.A., and Carrillo, L. (2025). COVID-19 and MPXV: Twindemic Response and Dual Infections in Individuals in a US Metro. Epidemiologia, 7.","DOI":"10.3390\/epidemiologia7010003"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Harapan, H., Ophinni, Y., Megawati, D., Frediansyah, A., Mamada, S.S., Salampe, M., Bin Emran, T., Winardi, W., Fathima, R., and Sirinam, S. (2022). Monkeypox: A Comprehensive Review. Viruses, 14.","DOI":"10.3390\/v14102155"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1007\/s00253-024-13361-6","article-title":"Monkeypox virus A29L protein as the target for specific diagnosis and serological analysis","volume":"108","author":"Liang","year":"2024","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/j.virol.2014.06.039","article-title":"A highly specific monoclonal antibody against monkeypox virus detects the heparin binding domain of A27","volume":"464\u2013465","author":"Hughes","year":"2014","journal-title":"Virology"},{"key":"ref_7","first-page":"452","article-title":"[Monkeypox]","volume":"77","author":"Absil","year":"2022","journal-title":"Rev. Med. Liege"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1378","DOI":"10.1016\/j.jiph.2022.11.002","article-title":"In-silico primer designing and PCR for detection of monkeypox virus (MPXV)","volume":"15","author":"Khoo","year":"2022","journal-title":"J. Infect. Public Health"},{"key":"ref_9","first-page":"69","article-title":"Emerging strategies for monkeypox: Antigen and antibody applications in diagnostics, vaccines, and treatments","volume":"12","author":"Wang","year":"2025","journal-title":"Mil. Med. Res."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Davis, I., Payne, J.M., Olguin, V.L., Sanders, M.P., Clements, T., Stefan, C.P., Williams, J.A., Hooper, J.W., Huggins, J.W., and Mucker, E.M. (2023). Development of a specific MPXV antigen detection immunodiagnostic assay. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1243523"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Qu, J., Zhang, X., Liu, K., Li, Y., Wang, T., Fang, Z., Chen, C., Tan, X., Lin, Y., and Xu, Q. (2024). A Comparative Evaluation of Three Diagnostic Assays for the Detection of Human Monkeypox. Viruses, 16.","DOI":"10.3390\/v16081286"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"10064","DOI":"10.1021\/acs.analchem.4c01802","article-title":"Sensitive Dual-Signal ELISA Based on Specific Phage-Displayed Double Peptide Probes with Internal Filtering Effect to Assay Monkeypox Virus Antigen","volume":"96","author":"Pang","year":"2024","journal-title":"Anal. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1080\/19420862.2016.1212149","article-title":"Phage display-derived human antibodies in clinical development and therapy","volume":"8","author":"Frenzel","year":"2016","journal-title":"mAbs"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.imlet.2015.07.005","article-title":"A new expression vector facilitating production and functional analysis of scFv antibody fragments selected from Tomlinson I+J phagemid libraries","volume":"167","author":"Ossysek","year":"2015","journal-title":"Immunol. Lett."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1007\/s13238-019-0639-7","article-title":"Phage display screening of therapeutic peptide for cancer targeting and therapy","volume":"10","author":"Saw","year":"2019","journal-title":"Protein Cell"},{"key":"ref_16","first-page":"1122","article-title":"Human monoclonal antibody reactive to stomach cancer produced by mouse-human hybridoma technique","volume":"77","author":"Yoshikawa","year":"1986","journal-title":"Jpn. J. Cancer Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"342514","DOI":"10.1016\/j.aca.2024.342514","article-title":"A rapid and sensitive fluorescent chromatography with cloud system for MPXV point-of-care diagnosis","volume":"1302","author":"Yan","year":"2024","journal-title":"Anal. Chim. Acta"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"100995","DOI":"10.1016\/j.jpha.2024.100995","article-title":"Optical biosensing of monkeypox virus using novel recombinant silica-binding proteins for site-directed antibody immobilization","volume":"14","author":"Song","year":"2024","journal-title":"J. Pharm. Anal."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e202400634","DOI":"10.1002\/biot.202400634","article-title":"Chicken-Derived Single-Chain Variable Fragments Targeting Monkeypox Virus A29L Protein","volume":"20","author":"Lee","year":"2025","journal-title":"Biotechnol. J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"113881","DOI":"10.1016\/j.jim.2025.113881","article-title":"Development of a one-step diagnostic and differential immunochromatographic method for mpox and chickenpox","volume":"541","author":"Li","year":"2025","journal-title":"J. Immunol. Methods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1039\/D2AN02043B","article-title":"Gold-based paper for antigen detection of monkeypox virus","volume":"148","author":"Ye","year":"2023","journal-title":"Analyst"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e0320025","DOI":"10.1128\/mbio.03200-25","article-title":"A cocktail vaccine with monkeypox virus antigens confers protection without selecting mutations in potential immune evasion genes in the vaccinia WR strain challenge","volume":"17","author":"Sun","year":"2026","journal-title":"mBio"}],"container-title":["Antibodies"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4468\/15\/3\/45\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T17:48:45Z","timestamp":1780076925000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4468\/15\/3\/45"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,29]]},"references-count":22,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["antib15030045"],"URL":"https:\/\/doi.org\/10.3390\/antib15030045","relation":{},"ISSN":["2073-4468"],"issn-type":[{"value":"2073-4468","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,29]]}}}