During screening for NANOBODY® molecules, large amounts of clones are tested for the desired binding and functional properties. As this is a labor-intensive step, the High-Throughput Profiling Unit at Ablynx has been expanding its multiplexing capacity in order to maximize the throughput and efficiency of the screening campaigns. A multiplex assay setup that allows for assessment of the binding of NANOBODY samples towards recombinant protein-coupled beads and target-expressing cells in the same well, was established. By implementing this workflow, our team is able to manage the ever-increasing demand in screening during early-stage projects and simultaneously assess cross-reactivity, target specificity and other biologically-relevant properties (e.g. ligand competition).