Wp2 - Selection of Monovalent Ligands


The employ of pre-screening and screening methods will be applied in order to reduce the number of potential ligands or multivalent systems, selecting the most active candidate ligands that will be used in in vitro studies by teams in WP5. Two type of selection test are envisaged which will be developed by two different groups: the Berlin and the Grenoble teams.

a. The Berlin group will use carbohydrate microarrays to test the affinity of different glycoconjugates for DC-SIGN. For this aim, the ligands will be adequately functionalized with long spacers and with a terminal amine that allows the attachment to a NHS-activated glass surface. The detection of DC-SIGN bound to the array will be performed using adequately fluorescence labelled antibodies. This will provide a first selection of those ligands.

b. The Grenoble team at CNRS-IBS will apply SPR techniques to screen monovalent ligands. Established techniques will be applied (inhibition of DC-SIGN binding to Man-BSA or other mannosylated proteins). This partner joint to Seville team, will develop of a new rapid screening method based on the use of a multivalent fluorescent probe to evaluate the ligands for DC-SIGN as monomers without any functionalization.

The tests carried out by the WP2 will provide a pool of compounds with good affinity for DC-SIGN and will reduce the number of multivalent systems to be prepared and the corresponding biological experiments to be performed as well.