| Autor: |
van Lith SA; Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands., van Duijnhoven SM; Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands., Navis AC; Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands., Leenders WP; Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands., Dolk E; QVQ Holding B.V. , Yalelaan 1, 3584 CL Utrecht, The Netherlands., Wennink JW; Department of Pharmaceutics, Utrecht University , Universiteitsweg 99, 3584 CG Utrecht, The Netherlands., van Nostrum CF; Department of Pharmaceutics, Utrecht University , Universiteitsweg 99, 3584 CG Utrecht, The Netherlands., van Hest JC; Department of Bio-organic Chemistry, Eindhoven University of Technology , P.O. Box 513 (STO 3.31), 5600 MB Eindhoven, The Netherlands. |
| Abstrakt: |
Conjugation of llama single domain antibody fragments (Variable Heavy chain domains of Heavy chain antibodies, VHHs) to diagnostic or therapeutic nanoparticles, peptides, proteins, or drugs offers many opportunities for optimized targeted cancer treatment. Currently, mostly nonspecific conjugation strategies or genetic fusions are used that may compromise VHH functionality. In this paper we present a versatile modular approach for bioorthogonal VHH modification and conjugation. First, sortase A mediated transPEGylation is used for introduction of a chemical click moiety. The resulting clickable VHHs are then used for conjugation to other groups employing the Cu + -independent strain-promoted alkyne-azide cycloadition (SPAAC) reaction. Using this approach, tail-to-tail bispecific VHHs and VHH-targeted nanoparticles are generated without affecting VHH functionality. Furthermore, this approach allows the bioconjugation of multiple moieties to VHHs for simple and convenient production of VHH-based theranostics. |
