Stech, M.M.StechRakotoarinoro, N.N.RakotoarinoroTeichmann, T.T.TeichmannZemella, A.A.ZemellaThoring, L.L.ThoringKubick, S.S.Kubick2022-03-062022-03-062021https://publica.fraunhofer.de/handle/publica/26853110.1007/978-1-0716-1406-8_9Cell-free protein synthesis (CFPS) enables the development of antibody conjugates, such as fluorophore conjugates and antibody-drug conjugates (ADCs), in a rapid and straightforward manner. In the first part, we describe the cell-free synthesis of antibodies containing fluorescent non-canonical amino acids (ncaa) by using pre-charged tRNA. In the second part, we describe the cell-free synthesis of antibodies containing ncaa by using an orthogonal system, followed by the site-specific conjugation of the fluorescent dye DyLight 650-phosphine. The expression of the antibodies containing ncaa was analyzed by SDS-PAGE, followed by autoradiography and the labeling by in-gel fluorescence. Two different fluorescently labeled antibodies could be generated.en610620book article