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In contrast to other hematologic malignancies, the development of effective antibody-based and cellular immunotherapies for AML has been limited by the scarcity of suitable target antigens.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We applied a phage display-based whole-cell panning method in which Fab-phage were biotinylated and captured, followed by next-generation sequencing (NGS), bioinformatic, and statistical analyses. Target deconvolution was performed using a CRISPR-Cas9 knockout library, fluorescence-activated cell sorting of antigen-negative cells, and NGS-based gRNA analysis. Selected candidates were further evaluated using primary AML patient cells and chimeric antigen receptor (CAR)-T cell assays.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We identified 28 unique monoclonal antibodies that preferentially bound AML cell lines. CRISPR-Cas9-based target deconvolution enabled efficient identification of three cognate antigens. Selected lead candidates were validated by staining primary cells from AML patients and were engineered into CAR constructs. CAR-T cells targeting the identified antigens mediated efficient eradication of AML cell lines and primary AML cells.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This integrated antibody-based antigen discovery and validation approach may accelerate the development of monoclonal antibody- and CAR-based immunotherapies for AML and other indications.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/abt\/tbag043","type":"journal-article","created":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T11:49:36Z","timestamp":1788954576000},"page":"576-594","source":"Crossref","is-referenced-by-count":0,"title":["Discovery of candidate antibodies and antigens for acute myeloid leukemia therapy by combining whole-cell phage display selection and CRISPR-Cas9 library 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