A Bioorthogonal Click Chemistry Toolbox for Targeted Synthesis of Branched and Well‐Defined Protein–Protein Conjugates. Issue 31 (26th May 2020)

• Record Type: Journal Article
• Authors: Baalmann, Mathis; Neises, Laura; Bitsch, Sebastian; Schneider, Hendrik; Deweid, Lukas; Werther, Philipp; Ilkenhans, Nadja; Wolfring, Martin; Ziegler, Michael J.; Wilhelm, Jonas; Kolmar, Harald; Wombacher, Richard
• Is Part Of: Angewandte Chemie international edition. Volume 59:Issue 31(2020)
• Page Start: 12885
• Page End: 12893
• Publication Date: 2020-05-26
• Subjects: antibody–drug conjugates -- bioorthogonal chemistry -- click chemistry -- protein ligation -- protein–protein conjugates; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201915079
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Bioorthogonal chemistry holds great potential to generate difficult‐to‐access protein–protein conjugate architectures. Current applications are hampered by challenging protein expression systems, slow conjugation chemistry, use of undesirable catalysts, or often do not result in quantitative product formation. Here we present a highly efficient technology for protein functionalization with commonly used bioorthogonal motifs for Diels–Alder cycloaddition with inverse electron demand (DAinv ). With the aim of precisely generating branched protein chimeras, we systematically assessed the reactivity, stability and side product formation of various bioorthogonal chemistries directly at the protein level. We demonstrate the efficiency and versatility of our conjugation platform using different functional proteins and the therapeutic antibody trastuzumab. This technology enables fast and routine access to tailored and hitherto inaccessible protein chimeras useful for a variety of scientific disciplines. We expect our work to substantially enhance antibody applications such as immunodetection and protein toxin‐based targeted cancer therapies. Abstract : A highly efficient method for protein functionalization with commonly used bioorthogonal motifs for Diels–Alder cycloaddition with inverse electron demand (DAinv ) is presented. This technology enables fast and routine access to tailored and hitherto inaccessible protein chimeras.