We report the creation of multivalent ligand surfaces for cell capture by conjugation of ligand-appended 2D peptide assemblies on an antifouling glass substrate. The sheet-like structures organize ligands into non-uniform, patchy patterns, enhancing multivalent cell targeting. A 155 % increase in captured cells was achieved compared to the presentation of the ligands on surfaces lacking the peptide sheets. Being orthogonal to the commonly used dendrimer- and cyclic organic molecular-based scaffolds, this peptide assembly-based approach offers a facile method to modulate the identity, number, and spatial distribution of ligands through controlled peptide coassembly. Using this method, we constructed a surface bearing two types of ligands, which demonstrates a 128 % enhancement in targeting selectivity between two model cells compared to the mono-ligand surface. These findings illustrate that integration of peptide assemblies into ligand substrates permits robust and effective manipulation of multivalent cell targeting, advancing the development of customizable cell-binding materials.