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**CD3-Folate** is a chemically defined bispecific conjugate consisting of an anti-CD3 Fab fragment linked to folate via an oxime bond using unnatural amino acid mutagenesis, designed to target cytotoxic T cells to folate receptor-positive (FR+) tumors. It binds CD3 on T cells and folate receptor alpha (FRα) on cancer cells, promoting T cell-mediated killing through rosette formation and cytotoxicity, with demonstrated efficacy in preclinical models of ovarian and nasopharyngeal cancers, including tumor elimination in xenograft mice even with non-activated PBMCs. Folate conjugation does not alter pharmacokinetics, maintaining a serum half-life of ~60 minutes, and limits off-target effects due to FRα's restricted expression in normal tissues.
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