Efficient, chemoselective synthesis of immunomicelles using single-domain antibodies with a C-terminal thioester |
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| Authors: | Sanne WA Reulen Ingrid van Baal Jos MH Raats Maarten Merkx |
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| Affiliation: | (1) Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, P.O. Box 513, 5600, MB, Eindhoven, the Netherlands;(2) Department of Biomolecular Chemistry, Radboud University Nijmegen, P.O. Box 9101, 6500, HB, Nijmegen, the Netherlands;(3) ModiQuest B.V., Heijendaalseweg 135, 6525, AJ, Nijmegen, the Netherlands |
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| Abstract: | Background Classical bioconjugation strategies for generating antibody-functionalized nanoparticles are non-specific and typically result in heterogeneous compounds that can be compromised in activity. Expression systems based on self-cleavable intein domains allow the generation of recombinant proteins with a C-terminal thioester, providing a unique handle for site-specific conjugation using native chemical ligation (NCL). However, current methods to generate antibody fragments with C-terminal thioesters require cumbersome refolding procedures, effectively preventing application of NCL for antibody-mediated targeting and molecular imaging. |
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