{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,21]],"date-time":"2025-11-21T12:24:31Z","timestamp":1763727871062,"version":"3.40.5"},"reference-count":40,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2020,12,13]],"date-time":"2020-12-13T00:00:00Z","timestamp":1607817600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vox Sanguinis"],"published-print":{"date-parts":[[2021,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background and objectives<\/jats:title><jats:p>Anti\u2010CD38 monoclonal antibodies, including daratumumab and isatuximab, often interfere with pretransfusion testing. Dithiothreitol (DTT) treatment of red blood cells (RBCs) negates this interference. However, the optimum DTT concentration and treatment time have not been well defined. Here, we quantified CD38 on RBCs before and after DTT treatment using a flow cytometric antibody binding assay (FABA) to specify the optimum conditions for CD38 inactivation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and methods<\/jats:title><jats:p>For FABA, untreated or DTT\u2010treated RBCs were incubated with fluorescein isothiocyanate\u2010labelled anti\u2010CD38 antibody, in the presence or absence of 100\u2010fold or more excess of unlabelled anti\u2010CD38 antibody, and then analysed by flow cytometry (FCM). Dissociation of CD38\u2010positive and control histograms was determined from the D\u2010value using the Kolmogorov\u2013Smirnov test. The results from FABA were compared with those from conventional FCM, indirect antiglobulin test (IAT) and Western blotting.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The results from FABA were more consistent than those from conventional FCM. The D\u2010value was found to be reliable in the analysis of difference between CD38 before and after DTT treatment. Our data showed that 0\u00b70075\u00a0mol\/l DTT for 30\u00a0min is sufficient to inactivate CD38 on RBCs. These results were stable and consistent with the findings from IAT.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Flow cytometric antibody binding assay is an objective way of evaluating the efficacy of DTT treatment for CD38 on RBCs. This approach allows the detection of a small number of cell surface antigens and will be useful for assessing the various chemical treatments to denature RBC antigens.<\/jats:p><\/jats:sec>","DOI":"10.1111\/vox.13052","type":"journal-article","created":{"date-parts":[[2020,12,13]],"date-time":"2020-12-13T12:43:22Z","timestamp":1607863402000},"page":"725-734","source":"Crossref","is-referenced-by-count":3,"title":["A newly devised flow cytometric antibody binding assay helps evaluation of dithiothreitol treatment for the inactivation of CD38 on red blood cells"],"prefix":"10.1111","volume":"116","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5202-1522","authenticated-orcid":false,"given":"Hiroaki","family":"Furumaki","sequence":"first","affiliation":[{"name":"Transfusion and Cell Therapy Hamamatsu University School of Medicine Hamamatsu 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