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The EuroFlow Consortium previously developed an 8\u2010color flow cytometric MRD protocol, effective for &gt;98% of BCP\u2010ALL patients treated with chemotherapy. This study aimed to enhance MRD detection, particularly for patients treated with CD19\u2010targeted therapies, by expanding the EuroFlow protocol to a 12\u2010color panel. This new panel incorporates additional B\u2010cell markers and exclusion T\/NK\u2010cell markers (CD3 and CD7). Through an evaluation of 237 diagnostic BCP\u2010ALL samples, CD22, CD24, and HLA\u2010DR were selected as additional B\u2010cell gating markers. Two 12\u2010color tubes were technically optimized and clinically validated across 101 patient follow\u2010up samples, demonstrating excellent concordance with molecular MRD levels (\n                    <jats:italic>R<\/jats:italic>\n                    <jats:sup>2<\/jats:sup>\n                    \u2009=\u20090.88). The 12\u2010color BCP\u2010ALL MRD tubes were compatible with the previously developed 8\u2010color automated gating and identification (AGI) tool and demonstrated good reproducibility. Our findings indicate that the 12\u2010color panel performs comparably to the 8\u2010color BCP\u2010ALL MRD panel, including both CD19\u2010positive and CD19\u2010negative cases. However, it offers an improved definition of the B\u2010cell lineage, particularly for expert\u2010guided manual data analysis, and provides additional information on the expression of the targetable marker CD22.\n                  <\/jats:p>","DOI":"10.1002\/hem3.70125","type":"journal-article","created":{"date-parts":[[2025,4,13]],"date-time":"2025-04-13T08:00:41Z","timestamp":1744531241000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Minimal residual disease assessment following CD19\u2010targeted therapy in B\u2010cell precursor acute lymphoblastic leukemia using standardized 12\u2010color flow cytometry: A EuroFlow study"],"prefix":"10.1002","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7240-5176","authenticated-orcid":false,"given":"Martijn W. 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