{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T02:06:24Z","timestamp":1747188384401,"version":"3.40.5"},"reference-count":24,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2023,11,12]],"date-time":"2023-11-12T00:00:00Z","timestamp":1699747200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion"],"published-print":{"date-parts":[[2023,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Antibodies against human neutrophil antigen (HNA) are involved in the pathogenesis of neonatal alloimmune neutropenia, autoimmune neutropenia, and transfusion\u2010related acute lung injury. The present methods for anti\u2010HNA antibody identification strongly depend on the presence of standard antisera with known allo\/isospecificities. Here, we aimed to produce recombinant humanized antibodies to HNA from available mouse monoclonal antibodies (MoAbs).<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design and Methods<\/jats:title><jats:p>RNAs were extracted from available hybridoma cells producing mouse anti\u2010HNA antibodies recognizing HNA\u20101a (TAG\u20101), \u20101b (TAG\u20102), \u20102 (TAG\u20104), and Fc\u03b3RIIIb, and the cDNA was synthesized. Recombinant fragments consisting of the variable regions of the H and L chains of the mouse MoAb ligated to the constant region of human IgG were incorporated into an expression vector and transfected into CHO cells. Antibody specificity of the selected humanized monoclonal antibodies was confirmed, and tested by the participants of the ISBT Granulocyte Immunobiology Working Party (GIWP) workshop 2020.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>GIFT results confirmed the specific reactivity of TAGH\u20101 to \u20104, except for a cross\u2010reactivity of TAGH\u20102 with HNA\u20101a\/a neutrophils, only in flow\u2010cytometry. MAIGA results showed clear specificity of all humanized antibodies, but the selection of the appropriate capture monoclonal antibody was essential for the test. The results of the ISBT GIWP showed high concordance among the labs.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>These are the first humanized monoclonal antibodies to HNA\u20101 and HNA\u20102 antigens produced and they will be important standard reagents for laboratories testing for neutrophil antibodies. We plan to have these humanized MoAbs available through WHO.<\/jats:p><\/jats:sec>","DOI":"10.1111\/trf.17590","type":"journal-article","created":{"date-parts":[[2023,11,12]],"date-time":"2023-11-12T18:43:42Z","timestamp":1699814622000},"page":"2282-2288","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Production of recombinant humanized monoclonal anti\u2010human neutrophil antigen (<scp>HNA<\/scp>) antibodies with potential applicability as standard antibodies"],"prefix":"10.1111","volume":"63","author":[{"given":"Yuko","family":"Ishimoto","sequence":"first","affiliation":[{"name":"Kanto\u2010Koshinetsu Block Blood Center Japanese Red Cross Society  Tokyo Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kikuyo","family":"Taniguchi","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Science Sanyo Women's College  Hiroshima 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