title: Legomedicine-A Versatile Chemo-Enzymatic Approach for the Preparation of Targeted Dual-Labeled Llama Antibody-Nanoparticle Conjugates.
abstract: 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.
authors.affiliation: Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.
authors.forename: Sanne A M
authors.initials: SA
authors.lastname: van Lith
authors.affiliation: Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.
authors.forename: Sander M J
authors.initials: SM
authors.lastname: van Duijnhoven
authors.affiliation: Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.
authors.forename: Anna C
authors.identifier: 0000-0002-5267-4678
authors.initials: AC
authors.lastname: Navis
authors.affiliation: Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.
authors.forename: William P J
authors.identifier: 0000-0003-0066-220X
authors.initials: WP
authors.lastname: Leenders
authors.affiliation: QVQ Holding B.V. , Yalelaan 1, 3584 CL Utrecht, The Netherlands.
authors.forename: Edward
authors.initials: E
authors.lastname: Dolk
authors.affiliation: Department of Pharmaceutics, Utrecht University , Universiteitsweg 99, 3584 CG Utrecht, The Netherlands.
authors.forename: Jos W H
authors.initials: JW
authors.lastname: Wennink
authors.affiliation: Department of Pharmaceutics, Utrecht University , Universiteitsweg 99, 3584 CG Utrecht, The Netherlands.
authors.forename: Cornelus F
authors.initials: CF
authors.lastname: van Nostrum
authors.affiliation: Department of Bio-organic Chemistry, Eindhoven University of Technology , P.O. Box 513 (STO 3.31), 5600 MB Eindhoven, The Netherlands.
authors.forename: Jan C M
authors.identifier: 0000-0001-7973-2404
authors.initials: JC
authors.lastname: van Hest
mesh_terms: D000480:Alkynes / Q000737:chemistry; D019881:Aminoacyltransferases / Q000378:metabolism; D000818:Animals; D001386:Azides / Q000737:chemistry; D001426:Bacterial Proteins / Q000378:metabolism; D002161:Camelids, New World / Q000276:immunology*; D057930:Click Chemistry / Q000379:methods; D061565:Cycloaddition Reaction / Q000379:methods; D003546:Cysteine Endopeptidases / Q000378:metabolism; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000378:metabolism; D061905:Single-Domain Antibodies / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism
chemical_list: D000480:Alkynes; D001386:Azides; D001426:Bacterial Proteins; D018796:Immunoconjugates; D007143:Immunoglobulin Heavy Chains; D061905:Single-Domain Antibodies; D011092:Polyethylene Glycols; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases