Rational design of an DNA-scaffolded high-affinity binder for langerin

Binders of Langerin could target vaccines to Langerhans cells for improved therapeutic effect. As Langerin has only low affinity for monovalent glycan ligands, highly multivalent presentation has previously been key for targeting. Aiming to reduce the amount of ligands required, we rationally designed molecularly defined high affinity binders based on the precise display of glycomimetic ligands (Glc2NTs) on DNA-PNA scaffolds. Rather than mimicking Langerin?s homotrimeric structure with a C3-symmetrical scaffold, we devised a strategy to improve readily accessible, easy to design bivalent binders. The method considers the requirements for bridging sugar binding sites and statistical rebinding as a means to both strengthen the interactions at single binding sites and amplify the avidity enhancement provided by chelation. The method enabled a 1150-fold net improvement over the affinity of the free ligand and provided a nanomolar binder (IC50 = 300 nM) for specific internalization by Langerin expressing cells

The Application of Dual‐Layer, Mussel‐Inspired, Antifouling Polyglycerol‐Based Coatings in Ventricular Assist Devices

Continuous‐flow ventricular assist devices (VADs) have established themselves as a lifesaving therapy option in patients with severe cardiovascular disease. Unfortunately, complications with VADs resulting from the shear‐induced formation of surface blood clots are common. In the current work, an antifouling coating based on the combination of mussel‐inspired dendritic polyglycerol (MI‐dPG) and linear polyglycerol (lPG) is tested for its cell‐repelling properties, biocompatibility, and complement activating properties. Furthermore, the adhesion and activation of blood platelets are tested under static and flow conditions. The adhesion and proliferation of two cell types are studied by means of LIVE/DEAD cell staining, and it is clearly observed that the lPG‐functionalized MI‐dPG coating prevents cell adhesion. Additionally, no cell mortality is observed on all substrates, indicating the biocompatibility of the tested coatings. All coatings show lower (or equal) complement‐activating properties than bare titanium, which is considered a highly biocompatible material. Most importantly, the lPG‐functionalized system prevents the adhesion and activation of blood platelets under static and flow conditions. Finally, a prototype VAD is successfully coated with MI‐dPG under flow conditions. In the current study, the efficient lPG‐functionalization of the MI‐dPG coating is proved to obtain cell‐ and platelet‐repelling surfaces

Rational Design of a DNA‐Scaffolded High‐Affinity Binder for Langerin

Binders of langerin could target vaccines to Langerhans cells for improved therapeutic effect. Since langerin has low affinity for monovalent glycan ligands, highly multivalent presentation has previously been key for targeting. Aiming to reduce the amount of ligand required, we rationally designed molecularly defined high‐affinity binders based on the precise display of glycomimetic ligands (Glc2NTs) on DNA‐PNA scaffolds. Rather than mimicking langerin's homotrimeric structure with a C3‐symmetric scaffold, we developed readily accessible, easy‐to‐design bivalent binders. The method considers the requirements for bridging sugar binding sites and statistical rebinding as a means to both strengthen the interactions at single binding sites and amplify the avidity enhancement provided by chelation. This gave a 1150‐fold net improvement over the affinity of the free ligand and provided a nanomolar binder (IC50=300 nM) for specific internalization by langerin‐expressing cells.Peer Reviewe