Figure 1.

Concept overview

Identification and expression of tumor-specific TCR was followed by chicken immunization. Afterward, splenic RNA was isolated, and cDNA was synthesized. Genes encoding the variable antibody domains were amplified and assembled as scFv gene string for yeast surface display and subsequent screening by FACS. TCR-binding scFvs were cloned into bacterial and mammalian vectors for scFv and scFv-Fc expression, respectively. After selection of a lead candidate, the TCR idiotype-specific binder was reformatted as full-length antibody and subsequently characterized. Generation of ADCs by antibody MMAE conjugation was conducted by two different approaches resulting in DARs of 2 and 2–8, respectively. Created with BioRender.com.