{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T23:34:09Z","timestamp":1793230449184,"version":"4.5.0"},"reference-count":37,"publisher":"SAGE Publications","issue":"6","license":[{"start":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T00:00:00Z","timestamp":1787788800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"name":"National Institute for Occupational Safety and Health","award":["OH012524"],"award-info":[{"award-number":["OH012524"]}],"id":[{"id":"https:\/\/ror.org\/0502a2655","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Institute for Occupational Safety and Health","award":["OH012726"],"award-info":[{"award-number":["OH012726"]}],"id":[{"id":"https:\/\/ror.org\/0502a2655","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Monoclonal Antibodies in Immunodiagnosis &amp; Immunotherapy"],"published-print":{"date-parts":[[2026,12]]},"abstract":"<jats:p>Methylene-diphenyl diisocyanate (MDI) and hexamethylene diisocyanate (HDI), important chemicals for polyurethane production, can react with self-molecules, trigger immune responses, and cause asthma. To validate molecular determinants of diisocyanate immune recognition and develop renewable immunoassay reagents, we generated MDI- and HDI-specific recombinant monoclonal antibodies (mAbs) from hybridoma-derived variable-region DNA sequences using a human embryonic kidney (HEK) cell line expression system. Three anti-MDI and one anti-HDI mouse IgG1\/\u03ba mAbs were produced, purified, and evaluated for binding to MDI, HDI, and control antigens, including toluene diisocyanate (TDI) and mock-reacted protein. To support assay standardization in research and translational applications, we engineered one anti-MDI (DA5) and one anti-HDI (AG5) mAb as human Fc chimeras (human IgG1 or IgE heavy-chain constant regions with human \u03ba light-chain constant region) and confirmed that chimerization preserved antigen specificity. As a proof-of-principle, one of the chimeric mAbs was evaluated as a candidate calibrator in an enzyme-linked immunosorbant assay (ELISA) format to quantify diisocyanate-specific antibodies in serum from an MDI-exposed worker. In addition, mAb DA5 was engineered with a FLAG epitope to enable immunoassays for detecting MDI\u2013protein reaction products in airway fluid, including sandwich ELISA (with anti-MDI mAb CE2) and Western blot formats. Together, these data show that diisocyanate specificity can be recapitulated by recombinant expression from deposited variable-region DNA sequences. The engineered reagents enable immunoassay detection of MDI\u2013protein adducts in complex matrices and serve as proof-of-concept calibrators for ELISA-based quantification of anti-diisocyanate antibodies in human serum.<\/jats:p>","DOI":"10.1177\/21679436261483330","type":"journal-article","created":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T13:14:31Z","timestamp":1787836471000},"page":"104-111","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":0,"title":["Recombinant Expression and Engineering of MDI- and HDI-Specific Monoclonal Antibodies for Proof-of-Concept Detection of Diisocyanate Adducts and Candidate Assay Calibrators"],"prefix":"10.1177","volume":"45","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2461-9409","authenticated-orcid":false,"given":"Adam V.","family":"Wisnewski","sequence":"first","affiliation":[{"name":"Department of Internal Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carrie A.","family":"Redlich","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Yale University, New Haven, Connecticut, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2026,8,27]]},"reference":[{"key":"e_1_3_2_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/pi.6441"},{"key":"e_1_3_2_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/pat.5277"},{"key":"e_1_3_2_4_1","unstructured":"3.U. S. Department of Veterans Affairs. CARC paint. 2026. Available from: https:\/\/www.publichealth.va.gov\/exposures\/carc-paint\/index.asp [Last accessed: April29 2026]."},{"key":"e_1_3_2_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.eurpolymj.2024.113421"},{"key":"e_1_3_2_6_1","volume-title":"Chemistry and Technology of Isocyanates","author":"Ulrich H","year":"1996","unstructured":"5.Ulrich H. Chemistry and Technology of Isocyanates. Wiley: New York; 1996."},{"key":"e_1_3_2_7_1","volume-title":"Hunstman and BASF put final touches on MDI projects","author":"Tullo A","year":"2022","unstructured":"6.Tullo A. Hunstman and BASF put final touches on MDI projects. C&EN Global Enterprise; 2022. Available from: https:\/\/cen.acs.org\/materials\/polymers\/Huntsman-BASF-put-final-touches\/100\/i26 [Last accessed: August20, 2026]"},{"key":"e_1_3_2_8_1","doi-asserted-by":"publisher","DOI":"10.1080\/15298669391355206"},{"key":"e_1_3_2_9_1","doi-asserted-by":"publisher","DOI":"10.3109\/9781841849256.018"},{"key":"e_1_3_2_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxrep.2025.102150"},{"key":"e_1_3_2_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajim.23323"},{"key":"e_1_3_2_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajim.22815"},{"key":"e_1_3_2_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajim.21010"},{"key":"e_1_3_2_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2011.03.007"},{"key":"e_1_3_2_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/0091-6749(93)90342-D"},{"key":"e_1_3_2_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1567-5769(01)00174-6"},{"key":"e_1_3_2_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/ftox.2022.836567"},{"key":"e_1_3_2_18_1","doi-asserted-by":"publisher","DOI":"10.1097\/00130832-200104000-00009"},{"key":"e_1_3_2_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbi.2026.111912"},{"key":"e_1_3_2_20_1","doi-asserted-by":"publisher","DOI":"10.1080\/10473220301441"},{"key":"e_1_3_2_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.toxlet.2014.12.012"},{"key":"e_1_3_2_22_1","doi-asserted-by":"publisher","DOI":"10.1080\/15459620601133779"},{"issue":"8","key":"e_1_3_2_23_1","first-page":"901","article-title":"Biomonitoring Hexamethylene diisocyanate (HDI) exposure based on serum levels of HDI-specific IgG","volume":"56","author":"Wisnewski AV","year":"2012","unstructured":"22.Wisnewski AV, , Stowe MH, , Nerlinger A, et al. Biomonitoring Hexamethylene diisocyanate (HDI) exposure based on serum levels of HDI-specific IgG. Ann Occup Hyg 2012;56(8):901\u2013910.","journal-title":"Ann Occup Hyg"},{"key":"e_1_3_2_24_1","doi-asserted-by":"publisher","DOI":"10.1093\/annweh\/wxab021"},{"key":"e_1_3_2_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-025-11730-3"},{"key":"e_1_3_2_26_1","doi-asserted-by":"publisher","DOI":"10.1089\/mab.2020.0006"},{"key":"e_1_3_2_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molimm.2012.11.017"},{"key":"e_1_3_2_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2013.05.022"},{"key":"e_1_3_2_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2016.02.005"},{"key":"e_1_3_2_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2010.01.037"},{"key":"e_1_3_2_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2011.08.011"},{"key":"e_1_3_2_32_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0343833"},{"key":"e_1_3_2_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(75)90431-5"},{"key":"e_1_3_2_34_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.chemrestox.8b00262"},{"key":"e_1_3_2_35_1","doi-asserted-by":"publisher","DOI":"10.1186\/1745-6673-6-6"},{"key":"e_1_3_2_36_1","doi-asserted-by":"publisher","DOI":"10.1093\/carcin\/16.10.2583"},{"key":"e_1_3_2_37_1","doi-asserted-by":"publisher","DOI":"10.1021\/tx5005002"},{"key":"e_1_3_2_38_1","doi-asserted-by":"publisher","DOI":"10.1021\/tx00005a005"}],"container-title":["Monoclonal Antibodies in Immunodiagnosis &amp; Immunotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/21679436261483330","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/full-xml\/10.1177\/21679436261483330","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/21679436261483330","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T23:17:26Z","timestamp":1793229446000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.sagepub.com\/doi\/10.1177\/21679436261483330"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,27]]},"references-count":37,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,12]]}},"alternative-id":["10.1177\/21679436261483330"],"URL":"https:\/\/doi.org\/10.1177\/21679436261483330","relation":{},"ISSN":["2167-9436","2167-9436"],"issn-type":[{"value":"2167-9436","type":"print"},{"value":"2167-9436","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,27]]}}}