To validate the ability of AS2 to induce ternary complex formation, we turned to size exclusion chromatography (SEC) as an orthogonal method to the previously performed native gel electrophoresis assay. Using CM11 as benchmark, we confirmed the formation of a VCB2:AS2 ternary complex, which migrated quicker than the VCB vehicle control (Figure ). Various conditions were tested, both with and without an excess of AS2 (green and lilac lines, Figure ), resulting in the complete or partial formation of the VCB2:AS2 ternary complex, respectively. This was confirmed by the retention time comparison observed for the CM11-mediated ternary complex. As controls, cisAS1 did not form any ternary complex, leading to identical retention time as VCB in vehicle as control. Together the results from both native gels and SEC indicate that AS2 works by effectively promoting the formation of a ternary complex, further supporting the general PROTAC-like behaviour of FerroTAC compounds.