Project Details
Projekt
Semisynthesis of antibody drug conjugates via biocatalysis
Applicant
Professor Dr. Frank Bordusa
Subject Area
Biological and Biomimetic Chemistry
Term
from 2012 to 2021
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 223395898
The development of novel and robust bioconjugation techniques for efficient and site-specific modification of proteins represents a considerable field in contemporary research. Beside the applicability of modified proteins for functional analysis in basic research, the conjugation of reporter molecules, polymers or drugs to antibodies within the scope of diagnostic or therapeutic applications possesses a high economic interest. Especially antibody drug conjugates (ADCs) evolved to an important tool regarding the treatment of cancer. Despite the high demand for novel ADCs, efficient modification methods for economical production of stable and functional conjugated antibodies in industrial scale are still rare. Particularly the synthesis of site-specifically modified antibodies to obtain homogeneous products constitutes an issue. Therefore, we are developing a universal biocatalysis platform for C-terminal derivatization of pharmaceutically relevant antibodies and antibody fragments based on a trypsin variant called trypsiligase. During the first three years of funding, we focused on the optimization of this biocatalyst by using a directed evolution approach. Based on the outstanding results, which are presented in this proposal, we intend to further improve this enzyme for site-specific modification of antibodies. Nevertheless, regarding the second funding period we will focus on the practical application of improved trypsiligase variants. Therefore we will modify a set of Fab fragments using several moieties and optimized trypsiligase variants. The resulting Fab conjugates will be analyzed regarding antigen binding, stability and cytotoxicity. Functionality in vivo will be examined by internalization experiments and the application to a mouse model.
DFG Programme
Priority Programmes
