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Off-target toxicity is a major cause of dose-limiting toxicity for antibodydrug conjugates (ADCs), mechanisms of which remain poorly understood. Here, we demonstrate that cytoskeleton-associated protein 5 (CKAP5) serves as a cell surface target for T-DM1 and that binding of T-DM1 to CKAP5 is mediated by payload (DM1). This study introduces a novel molecular mechanism of ADC payload-mediated interaction with cell surface molecules to induce cytotoxicity. Upon binding to CKAP5, T-DM1 causes celldoi:10.18632/oncotarget.26461 fatcat:izsyzodzqbcl7b6mummzzfczje