{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T22:05:54Z","timestamp":1792101954214,"version":"4.4.0"},"reference-count":53,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2024,2,24]],"date-time":"2024-02-24T00:00:00Z","timestamp":1708732800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/100009425","name":"Landsteiner Foundation for Blood Transfusion Research","doi-asserted-by":"publisher","award":["1908"],"award-info":[{"award-number":["1908"]}],"id":[{"id":"10.13039\/100009425","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion"],"published-print":{"date-parts":[[2024,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a condition during pregnancy, which can lead to thrombocytopenia and a bleeding tendency with intracranial hemorrhage (ICH) being the most concerning complication in the fetus or neonate. An incompatibility between human platelet antigen (HPA)\u20101a accounts for the majority of FNAIT cases. Binding of HPA\u20101a\u2010specific alloantibodies to their target on fetal platelets and endothelial cells can induce apoptosis of megakaryocytes, disrupt platelet function, and impair angiogenesis. Currently, there is no screening program to identify pregnancies at risk for severe disease. A better understanding of HPA\u20101a\u2010specific antibody heterogeneity in FNAIT could aid in identifying pathogenic antibody properties linked to severe disease.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design and Methods<\/jats:title><jats:p>This study aimed to isolate HPA\u20101a\u2010specific B\u2010cells from an HPA\u20101a\u2010alloimmunized pregnant woman. Using fluorescently labeled HPA\u20101a\u2010positive platelets, single B\u2010cells were sorted and cultured for 10\u2009days to stimulate antibody production. Subsequently, supernatants were tested for the presence of antibodies by enzyme\u2010linked immunosorbent assay and their reactivity towards HPA\u20101a\u2010positive platelets. Amplification and sequencing of variable regions allowed the generation of monoclonal antibodies using a HEK\u2010Freestyle\u2010based expression system.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Three platelet\u2010specific B\u2010cells were obtained and cloned of which two were specific for HPA\u20101a, named D\u2010 and M\u2010204, while the third was specific for HLA class I, which was named L\u2010204.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>This study outlined an effective method for the isolation of HPA\u20101a\u2010specific B\u2010cells and the generation of monoclonal antibodies. Further characterization of these antibodies holds promise for better understanding the pathogenic nature of alloantibodies in FNAIT.<\/jats:p><\/jats:sec>","DOI":"10.1111\/trf.17758","type":"journal-article","created":{"date-parts":[[2024,2,24]],"date-time":"2024-02-24T08:39:37Z","timestamp":1708763977000},"page":"893-905","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Generation of human antibodies targeting human platelet antigen (<scp>HPA)<\/scp>\u20101a"],"prefix":"10.1111","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6749-2594","authenticated-orcid":false,"given":"Janita J.","family":"Oosterhoff","sequence":"first","affiliation":[{"name":"Department of Experimental Immunohematology Sanquin Research  Amsterdam The Netherlands"},{"name":"Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research 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