{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T04:12:21Z","timestamp":1778904741193,"version":"3.51.4"},"reference-count":40,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T00:00:00Z","timestamp":1772755200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T00:00:00Z","timestamp":1772755200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100012399","name":"Biomedical Advanced Research and Development Authority","doi-asserted-by":"publisher","award":["#75A50123C00052"],"award-info":[{"award-number":["#75A50123C00052"]}],"id":[{"id":"10.13039\/100012399","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Transfusion"],"published-print":{"date-parts":[[2026,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      Twenty\u2010four\u2010hour post\u2010transfusion recovery (PTR24) is a key measure of red blood cell (RBC) quality. Traditionally assessed using radioactive chromium\u201051 (\n                      <jats:sup>51<\/jats:sup>\n                      Cr), supply and facility limitations have prompted the need for alternatives. Biotinylation of RBCs (BioRBCs) offers a non\u2010radioactive method capable of distinguishing multiple RBC populations by flow cytometry. Researchers at three major blood centers developed a standardized BioRBC labeling protocol for pharmacokinetic studies.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Study Design and Methods<\/jats:title>\n                    <jats:p>The multicenter development of the standardized protocol followed a two\u2010phase design. First, inter\u2010 and intra\u2010site flow cytometry variability was assessed using RBCs biotinylated at 6 and 18\u2009\u03bcg\/mL. Samples were stained with streptavidin\u2010phycoerythrin (SA\u2010PE), diluted, and analyzed in triplicates using shared flow cytometry templates. Second, inter\u2010site variability of biotinylation was tested using paired RBC units split and processed simultaneously at each site. Separation index (SI) was calculated as: SI\u2009=\u2009(median BioRBC\u2009\u2212 median unlabeled)\/(84th percentile unlabeled\u2009\u2212\u2009median unlabeled)\/0.995.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Staining and analysis of BioRBCs demonstrated minimal inter\u2010 and intra\u2010site variability when using identical protocols. Coefficients of variation for median fluorescence intensity were &lt;7.9% within sites and &lt;5.1% across sites. Biotinylation of paired RBC with 6 and 18\u2009\u03bcg\/mL of biotin resulted in comparable separation indices, suggesting reliable and reproducible biotinylation ratios.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      The standardized BioRBC protocol produces reliable, reproducible labeling with minimal inter\u2010site variability. In vivo performance will be assessed in upcoming PTR24 studies comparing BioRBCs with dual radiolabeling using\n                      <jats:sup>51<\/jats:sup>\n                      Cr and technetium\u201099m (\n                      <jats:sup>99m<\/jats:sup>\n                      Tc).\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/trf.70157","type":"journal-article","created":{"date-parts":[[2026,3,8]],"date-time":"2026-03-08T20:31:37Z","timestamp":1773001897000},"page":"923-932","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Multi\u2010center development and validation of a standardized protocol for biotinylated red blood cell analysis"],"prefix":"10.1111","volume":"66","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7862-6201","authenticated-orcid":false,"given":"Susanne","family":"Marschner","sequence":"first","affiliation":[{"name":"Vitalant Research Institute  Denver 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