Integrated scFv identification and CAR T cell generation for AML targeting in vivo

Cancer immunotherapy has witnessed remarkable advancements, particularly in the development of chimeric antigen receptor (CAR) T cell therapy. Here, we integrated single chain variable fragment (scFv) development with CAR T cell generation based on a newly developed scFv phagemid library. High‐throughput long‐read PacBio sequencing identified 4.5 × 10 7 unique full‐length scFv proteins within the generated library. As a proof of principle, we screened for scFvs targeting C‐type lectin‐like molecule‐1 (CLL1) with subsequent cloning into a third generation retroviral CAR backbone. Functional assays revealed the specificity and potency of these CAR T cells in targeting CLL1‐positive AML cells in vitro. In vivo studies reduced tumor burden and improved survival rates compared to controls. Taken together, screening for tumor‐specific scFvs against CLL1 can rapidly generate AML‐specific CAR T cells with effective tumor killing in vivo.