van Lith Sanne A M, van Duijnhoven Sander M J, Navis Anna C, Leenders William P J, Dolk Edward, Wennink Jos W H, van Nostrum Cornelus F, van Hest Jan C M
Department of Pathology, Radboud University Nijmegen Medical Centre , Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.
QVQ Holding B.V. , Yalelaan 1, 3584 CL Utrecht, The Netherlands.
Bioconjug Chem. 2017 Feb 15;28(2):539-548. doi: 10.1021/acs.bioconjchem.6b00638. Epub 2017 Jan 18.
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.
将羊驼单域抗体片段（重链抗体的可变重链结构域，VHHs）与诊断或治疗性纳米颗粒、肽、蛋白质或药物进行偶联，为优化靶向癌症治疗提供了诸多机会。目前，大多使用的是非特异性偶联策略或基因融合方法，这可能会损害VHH的功能。在本文中，我们提出了一种用于生物正交VHH修饰和偶联的通用模块化方法。首先，分选酶A介导的转聚乙二醇化用于引入化学点击基团。然后，所得的可点击VHHs用于通过不依赖铜的应变促进炔-叠氮环加成（SPAAC）反应与其他基团偶联。使用这种方法，可以生成头对头双特异性VHHs和VHH靶向纳米颗粒，而不会影响VHH的功能。此外，这种方法允许将多个部分生物偶联到VHHs上，以便简单方便地生产基于VHH的诊疗试剂。
