Synthesis of Fluorescently Labeled Antibodies Using Non-Canonical Amino Acids in Eukaryotic Cell-Free Systems

Abstract Cell-free protein synthesis (CFPS) enables the development of antibody conjugates, such as fluorophore conjugates and antibody-drug (ADCs), in a rapid straightforward manner. In first part, we describe cell-free antibodies containing fluorescent non-canonical amino acids (ncaa) by using pre-charged tRNA. second ncaa an orthogonal system, followed site-specific conjugation dye DyLight 650-phosphine. The expression was analyzed SDS-PAGE, autoradiography labeling in-gel fluorescence. Two different fluorescently labeled could be generated.