Defining the property space for chromatographic ligands from a homologous series of mixed-mode ligands

•Novel ligands were created to identify structural variants with altered selectivity for proteins.•Hydrophobic interactions were affected by changing solvent exposure and nearby substituents.•pH gradients were used to demonstrate alternative selectivity of proteins on multimodal resins.•A chemometric model was created that could predict protein retention on several multimodal resins.