{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T12:39:11Z","timestamp":1791549551801,"version":"4.3.4"},"reference-count":36,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:00:00Z","timestamp":1759968000000},"content-version":"vor","delay-in-days":8,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2025,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Quantification of T\u2010cells, B\u2010cells, and NK\u2010cells assay is crucial for diagnosing and monitoring immune diseases and evaluating lymphodepleting therapies. To standardize and validate an optimized workflow of the Beckman Coulter DuraClone IM Phenotyping Basic Kit for quantification of TBNK subsets in peripheral blood. Procedural changes included the use of an alternative lysis buffer, the addition of counting beads, the elimination of centrifugation steps, and an increase in acquisition volume. Validation followed CLSI H42\u2010A2 and H62 guidelines, assessing accuracy, precision, linearity, and limit of quantification (LLOQ). Accuracy was evaluated by comparison with Immuno\u2010Trol controls and by Bland\u2013Altman analysis against standard immunophenotyping methods. External proficiency was assessed through participation in the College of American Pathologists (CAP) TBNK program in 2024. Procedural steps were reduced by 50%, and processing time by 38.6%. The modified method demonstrated high accuracy (\u22123 &lt;\u2009bias &lt;\u200935; cells\/\u03bcL), a low bias based on Immuno\u2010Trol targets, and strong agreement in the Bland\u2013Altman analysis. The method successfully passed three CAP external proficiency tests in 2024, confirming interlaboratory reliability. Coefficients of variation for precision were below 10% for all subsets. Linearity exceeded\n                    <jats:italic>R<\/jats:italic>\n                    <jats:sup>2<\/jats:sup>\n                    \u2009&gt;\u20090.99 across clinically relevant ranges. Most subsets demonstrated an LLOQ below 10\u201350\u2009cells\/\u03bcL, which is suitable for clinical applications. The proposed modifications to the DuraClone IM kit protocol improved workflow efficiency and analytical performance without compromising accuracy or reproducibility. The validated method provides a standardized, reliable, and time\u2010efficient alternative for lymphocyte subset quantification.\n                  <\/jats:p>","DOI":"10.1002\/cyto.a.24967","type":"journal-article","created":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T07:01:38Z","timestamp":1759993298000},"page":"695-708","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Validation of an Optimized\n                    <scp>DuraClone<\/scp>\n                    Phenotyping Kit Workflow for\n                    <scp>TBNK<\/scp>\n                    Subset Quantification"],"prefix":"10.1002","volume":"107","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7478-0482","authenticated-orcid":false,"given":"Carlos Agustin","family":"Villegas\u2010Valverde","sequence":"first","affiliation":[{"name":"Abu Dhabi Stem Cells Center  Abu Dhabi UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0432-7943","authenticated-orcid":false,"given":"Imilla Casado","family":"Hernandez","sequence":"additional","affiliation":[{"name":"Abu Dhabi Stem Cells Center  Abu Dhabi UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9483-9971","authenticated-orcid":false,"given":"Yaima Zu\u00f1iga","family":"Rosales","sequence":"additional","affiliation":[{"name":"Abu Dhabi Stem Cells Center  Abu Dhabi UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-6699-1667","authenticated-orcid":false,"given":"Fatma","family":"Abdou","sequence":"additional","affiliation":[{"name":"Abu Dhabi Stem Cells Center  Abu Dhabi UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1109-3348","authenticated-orcid":false,"given":"Shadi Sharif","family":"Shamat","sequence":"additional","affiliation":[{"name":"Abu Dhabi Stem Cells Center  Abu Dhabi UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6209-0393","authenticated-orcid":false,"given":"Antonio Alfonso","family":"Bencomo\u2010Hernandez","sequence":"additional","affiliation":[{"name":"Abu Dhabi Stem Cells Center  Abu Dhabi UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5373-3414","authenticated-orcid":false,"given":"Yendry","family":"Ventura\u2010Carmenate","sequence":"additional","affiliation":[{"name":"Abu Dhabi Stem Cells Center  Abu Dhabi UAE"},{"name":"United Arab Emirates University\u2010 College of Medicine  Abu Dhabi UAE"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,10,9]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.21400"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/eji.202170126"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.22195"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.21930"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u20101\u20100716\u20102811\u20109_6"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells12141875"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.658314"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.23901"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20623"},{"key":"e_1_2_9_11_1","volume-title":"Assay Development and Validation of T, B, and NK Lymphocyte Subset Enumeration","author":"Illingworth A.","year":"2024"},{"key":"e_1_2_9_12_1","volume-title":"Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline\u2014Second Edition. 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