Evaluation of selectivity in homologous multimodal chromatographic systems using in silico designed antibody fragment libraries

•Fab variants were designed in silico and produced for chromatographic evaluations.•The CDR regions were shown to be an important binding site on most but not all Fabs.•Multimodal ligand design was important for separating different Fab variants.•Different ligands had distinct selectivity for separating various Fab variants.•Surface property analyses were valuable for evaluating chromatographic behavior.